Morphic forms of g1t38 and methods of manufacture thereof

ABSTRACT

This invention provides an unexpectedly stable, highly crystalline form of the di-HCl salt of 
     
       
         
         
             
             
         
       
     
     for advantageous therapeutic pharmaceutical efficacy and dosage form stability.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/721,631, filed Dec. 19, 2019, which is a continuation ofInternational Application No. PCT/US2018/040435, filed in theInternational Patent Cooperation Treaty, U.S. Receiving Office on Jun.29, 2018, which claims the benefit of and priority to U.S. ProvisionalApplication 62/526,937 which was filed on Jun. 29, 2017. The entirety ofeach of these applications is hereby incorporated by reference for allpurposes.

FIELD OF THE INVENTION

This invention provides an advantageous isolated morphic form of thedi-HCl salt, of G1T38, which is(2′-((5-(4-isopropylpiperazin-1-yl)pyridin-2-yl)amino)-7′,8′-dihydro-6′H-spiro[cyclohexane-1,9′-pyrazino[1′,2′:1,5]pyrrolo[2,3-d]pyrimidin]-6′-one).

BACKGROUND

U.S. Pat. Nos. 8,822,683; 8,598,197; 8,829,102 and 9,102,683 andcorresponding WO 2012/061156 assigned to G1 Therapeutics, Inc. describea class of N-(heteroaryl)-pyrrolo[3,2-d]pyrimidin-2-amine cyclindependent kinase inhibitors including2′-((5-(4-isopropylpiperazin-1-yl)pyridin-2-yl)amino)-7′,8′-dihydro-6′H-spiro[cyclohexane-1,9′-pyrazino[1′,2′:1,5]pyrrolo[2,3-d]pyrimidin]-6′-one(Compound 1) with the formula

The compound is currently referred to as “G1T38”. The di-HCl salt ofG1T38 (Compound 2) is currently in Phase Ib/2a human clinical trials inthe United States with the U.S. Food and Drug Administration for thetreatment of estrogen positive, HER2-negative breast cancer afterendocrine therapy failure. G1T38 has also been favorably evaluated in aPhase 1a toxicity trial in 75 women and found to be well tolerated withno significant adverse events.

G1T38 induces inhibition of cell proliferation in a variety ofCDK4/6-dependent tumorigenic cell lines including breast, melanoma,leukemia, and lymphoma cells and inhibits RB phosphorylation in vitroand in vivo. Additional favorable therapeutic properties of G1T38,including the selectivity for tumors over plasma in mouse xenografttumors, are highlighted in an article recently released in a peerreviewed journal (Bisi, et al., Preclinical development of G1T38: Anovel, potent and selective inhibitor of cyclin dependent kinases 4/6for use as an oral antineoplastic in patients with CDK 4/6 sensitivetumors”, Oncotarget, Mar. 15, 2017). See also U.S. Pat. No. 9,527,857.

Other publications that describe compounds of this general class includethe following. WO 2014/144326 filed by Strum et al. and assigned to G1Therapeutics describes compounds and methods for protection of normalcells during chemotherapy using pyrimidine based CDK4/6 inhibitors. WO2014/144596 filed by Strum et al. and assigned to G1 Therapeuticsdescribes compounds and methods for protection of hematopoietic stem andprogenitor cells against ionizing radiation using pyrimidine basedCDK4/6 inhibitors. WO 2014/144847 filed by Strum et al. and assigned toG1 Therapeutics describes HSPC-sparing treatments of abnormal cellularproliferation using pyrimidine based CDK4/6 inhibitors. WO2014/144740filed by Strum et al. and assigned to G1 Therapeutics describes highlyactive anti-neoplastic and anti-proliferative pyrimidine based CDK 4/6inhibitors. WO 2015/161285 filed by Strum et al. and assigned to G1Therapeutics describes tricyclic pyrimidine based CDK inhibitors for usein radioprotection. WO 2015/161287 filed by Strum et al. and assigned toG1 Therapeutics describes analogous tricyclic pyrimidine based CDKinhibitors for the protection of cells during chemotherapy. WO2015/161283 filed by Strum et al. and assigned to G1 Therapeuticsdescribes analogous tricyclic pyrimidine based CDK inhibitors for use inHSPC-sparing treatments of RB-positive abnormal cellular proliferation.WO 2015/161288 filed by Strum et al. and assigned to G1 Therapeuticsdescribes analogous tricyclic pyrimidine based CDK inhibitors for use asanti-neoplastic and anti-proliferative agents. WO 2016/040858 filed byStrum et al. and assigned to G1 Therapeutics describes the use ofcombinations of pyrimidine based CDK4/6 inhibitors with otheranti-neoplastic agents. WO 2016/040848 filed by Strum et al. andassigned to G1 Therapeutics describes compounds and methods for treatingcertain Rb-negative cancers with CDK4/6 inhibitors and topoisomeraseinhibitors.

Other biologically active fused spirolactams and their syntheses aredescribed, for example, in the following publications. Griffith, D. A.,et al. (2013). “Spirolactam-Based Acetyl-CoA Carboxylase Inhibitors:Toward Improved Metabolic Stability of a Chromanone Lead Structure.”Journal of Medicinal Chemistry 56(17): 7110-7119, describesmetabolically stable spirolactams wherein the lactam resides on thefused ring for the inhibition of acetyl-CoA carboxylase. WO 2013/169574filed by Bell et al. describes aliphatic spirolactams as CGRP receptorantagonists wherein the lactam resides on the spiro ring. WO 2007/061677filed by Bell et al. describes aryl spirolactams as CGRP receptorantagonists wherein the lactam resides on the spiro ring. WO 2008/073251filed by Bell et al. describes constrained spirolactam compounds whereinthe lactam resides on the spiro ring as CGRP receptor antagonists. WO2006/031606 filed by Bell et al. describes carboxamide spirolactamcompounds wherein the spirolactam resides on the spiro ring as CGRPreceptor antagonists. WO 2006/031610, WO 2006/031491, and WO 2006/029153filed by Bell et al. describe anilide spirolactam compounds wherein thespirolactam resides on the spiro ring. WO 2008/109464 filed by Bhunai etal. describes spirolactam compounds wherein the lactam resides on thespiro ring which is optionally further fused.

Given the therapeutic importance of G1T38 to patients suffering from aproliferative disorder such as a tumor or cancer, it would be beneficialto provide an advantageous means for delivery that may increasetherapeutic activity and/or stability.

SUMMARY

It has been discovered that Compound 2, di-HCl salt of G1T38(2′-((5-(4-isopropylpiperazin-1-yl)pyridin-2-yl)amino)-7′,8′-dihydro-6′H-spiro[cyclohexane-1,9′-pyrazino[1′,2′1,5]pyrrolo[2,3-d]pyrimidin]-6′-one)can be prepared in a highly purified, advantageous morphic form,referred to herein as Form B.

Form B of Compound 2 is an unexpected, highly stable, highly crystallineform of solid Compound 2, which is beneficial for therapeutic efficacyand for the manufacture of pharmaceutical formulations. As discussed inExample 4, Form B is stable under thermal stress of 60° C. for 7 days.Additionally, a long-term stability study at 25° C. and 60% relativehumidity revealed that isolated Compound 2 Form B is stable for at least1 year (Example 7). In one embodiment isolated Compound 2 Form B isstable for at least about 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, or24 months.

A number of crystallization and slurry experiments were conducted(Example 2, Tables 1-4) by varying temperature, cooling procedure, andisolation procedure. From these experiments, eleven unique forms ofCompound 2 were discovered, but only Form A, Form B, and Form D wereappropriate for evaluation. The other forms resulted in weak crystallineforms, solvates, unstable hydrates, or anhydrates. Of the three solidforms, Form B was discovered to be an unexpectedly superior highlycrystalline stable material for therapeutic dosage forms. In the dynamicvapor sorption experiment, Compound 2 remained in Form B after exposureto 90% relative humidity (Example 3).

Form B has advantageous properties for use as an active pharmaceuticalingredient in a solid dosage form and may have increased efficacy insuch a formulation. In one embodiment, Form B is produced byrecrystallization from HCl and acetone, as described in more detailbelow. In one embodiment, Form B is characterized by an XRPD patternsubstantially similar to that set forth in FIG. 7 . In one embodiment,Form B is characterized by an XRPD pattern comprising at least three2theta values selected from 6.5°±0.2, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°,18.1±0.2°, 19.7±0.2°, and 22.4±0.2°. In one embodiment, Form B ischaracterized by an XRPD pattern comprising at least the 2theta valuesof 9.5±0.2°. In some embodiments isolated Compound 2, Form B ischaracterized by the absence of at least one of the peaks at 4.6±0.2°2theta. In some embodiments isolated Compound 2, Form B is characterizedby the absence of a peak at 5.0±0.2° 2theta. In one embodiment, isolatedForm B is characterized as having a 7.5% weight loss between 31 and 120°C. in a thermogravimetric infrared (TG-IR) analysis. In one embodiment,isolated Form B is characterized as having differential scanningcalorimetry (DSC) onset endotherms at about 105±20° C., about 220±20°C., and about 350±20° C., for example at 105° C., 220° C., and 350° C.or 92° C., 219° C., and 341° C.

Thus, the present invention generally provides an isolated morphic FormB of Compound 2, pharmaceutical compositions containing such morphicform, methods of inhibiting or reducing the activity of CDK4 or CDK6 ina host using said isolated morphic form, and treating a host having apRb-positive cancer such as, for example, estrogen receptor-positive(ER+) breast cancer, non-small cell lung cancer (NSCLC), or prostatecancer, using the morphic form described herein, and methods ofpreparing such morphic form.

Compound 2 Form B can be produced, for example, by recrystallizingCompound 1 in concentrated HCl and acetone. In one embodiment, Compound1 is dissolved in concentrated HCl and heated. This is followed by theaddition of acetone and isolation of the product by cooling andfiltration.

In one embodiment, Compound 2 Form B is produced by therecrystallization of Compound 2 Form D. In an alternative embodiment,Compound 2 Form B is produced by repeated recrystallizations. In oneembodiment, pure Compound 2 Form B is purified from impure Compound 2Form B by a water:acetone (1:2) (v/v) slurry followed by vacuum drying.

Compound 2 Form A has less stability than Form B. Form A was producedwhen MeOH, EtOH, and 1-BuOH were used as solvents in the single solventcrystallizations and it was also produced in the binary solventcrystallizations using water and MeOH as the primary solvent. Slurryexperiments using n-heptane and c-hexane produced Form A as well.

Compound 2 Form D has less stability than Form B. In one embodiment,Form D is produced by stirring a slurry of Compound 2 in acetonitrile atroom temperature. In another embodiment, Form D is produced bydissolving Compound 1 in concentrated HCl before heating. Then thesolution is allowed to cool and acetone is only added aftercrystallization begins to drive the precipitation to completion. Theprecipitate is then isolated via filtration. In an alternativeembodiment, Form D is produced by dissolving Compound 1 in concentratedHCl before heating. Then the solution is allowed to cool and acetone isonly added once crystallization has occurred and all solids arecollected via filtration.

In alternative embodiments, a combination of two or more Forms ofCompound 2 is provided, such as Forms B and D; Forms B and A; or Forms Aand D. In an alternative embodiment, an isolated combination of threeforms is provided, for example, Forms A, B, and D.

In one embodiment a pharmaceutical composition is provided comprisingisolated Compound 2 morphic Form B and a pharmaceutically acceptableexcipient. In another embodiment, the pharmaceutical composition furthercomprises one or more additional therapeutic agents, for example but notlimited to, an anti-estrogen, anti-androgen, an antineoplastic agent, anaromatase inhibitor, a Bruton's tyrosine kinase (BTK) inhibitor, a CYP17inhibitor, an extracellular signal-regulated kinase (ERK) inhibitor, agonadotropin releasing hormone superagonist (GnRH agonist), aluteinizing hormone-releasing hormone (LH-RH) agonist, a luteinizinghormone-releasing hormone (LH-RH) antagonist, a mechanistic target ofrapamycin (mTOR) inhibitor, a mitogen-activated protein kinase (MEK)inhibitor, a nucleoside or nucleotide analogue or prodrug, aphosphatidylinositol 3-kinase (PI3K) pathway inhibitor, a rapidlyaccelerated fibrosarcoma (RAF) kinase inhibitor, a renin-angiotensinsystem (RAS) inhibitor, a selective estrogen receptor degrader (SERD), aselective estrogen receptor modulator (SERM), a serine-threonine proteinkinase B (Akt) inhibitor, or a topoisomerase inhibitor. In oneembodiment, the one or more additional therapeutic agents are selectedfrom letrazole, anastrozole, fulvestrant, tamoxifen, etoposide,enzalutamide, pictilisib, exemestane, or a combination thereof.

In another embodiment Compound 2 morphic Form B is used in combinationwith a SERD described in WO 2017/100712, WO 2017/100715, US2017/0166550, or US 2017/0166551. In yet another embodiment apharmaceutical composition is provided comprising isolated Compound 2morphic Form B, a pharmaceutically acceptable excipient, and a SERDdescribed in WO 2017/100712, WO 2017/100715, US 2017/0166550, or US2017/0166551.

In one aspect of the present invention, a method for treating a CDK4/6dependent cellular proliferation disorder is provided comprisingadministering to a host in need thereof a therapeutically effectiveamount of isolated Form B of Compound 2.

Also provided is the use of isolated morphic Form B in the manufactureof a medicament for treating a pRb-positive cancer, such as estrogenreceptor positive (ER+) breast cancer, non-small cell lung cancer(NSCLC), prostate cancer, or other abnormal cellular proliferation in ahost.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a comparison of XRPD patterns of Form A, Form B, and Form C.These three forms were obtained from crystallization and slurryexperiments as described in Example 2 and shown in Tables 1-4. Thex-axis is 2Theta measured in degrees and the y-axis is intensitymeasured in counts.

FIG. 2 is a comparison of XRPD patterns of Form D, Form E, and Form F.These three forms were obtained from crystallization and slurryexperiments as described in Example 2 and shown in Tables 1-4. Thex-axis is 2Theta measured in degrees and the y-axis is intensitymeasured in counts.

FIG. 3 is a comparison of XRPD patterns of Form G and Form H. These twoforms were obtained from crystallization and slurry experiments asdescribed in Example 2 and shown in Tables 1-4. Form G is an anhydrateand Form H is an n-PrOH solvate. The x-axis is 2Theta measured indegrees and the y-axis is intensity measured in counts.

FIG. 4A is a dynamic vapor sorption analysis showing the results from amoisture sorption experiment of Form A (Example 3). The material wasfound to be unstable and the XRPD analysis of dried sample at theconclusion of the experiment revealed a new Form, Form K. Form Aadsorbed 14.9 wt % at 60% RH (relative humidity) and 15.8 wt % at 90%RH. The x-axis is relative humidity measured as a percent and the y-axisis weight of water of the material measured as a percent.

FIG. 4B is a dynamic vapor sorption analysis showing the results from amoisture sorption experiment of Form D (Example 3). The material wasfound to be unstable and the XRPD analysis of dried sample at theconclusion of the experiment revealed a new Form, Form K. Form Dadsorbed 4.4 wt % at 60% RH (relative humidity) and 4.4 wt % at 90% RH.The x-axis is relative humidity measured as a percent and the y-axis isweight of water of the material measured as a percent.

FIG. 4C is a dynamic vapor sorption analysis showing the results from amoisture sorption experiment of Form B (Example 3). The material isstable and the XRPD analysis of a dried sample at the conclusion of theexperiment confirmed Form B. Form B adsorbed 5.8 wt % at 60% RH(relative humidity), and 5.9 wt % at 90% RH. The x-axis is relativehumidity measured as a percent and the y-axis is weight of water of thematerial measured as a percent.

FIG. 5A is a comparison of XRPD patterns of Form A before the moisturesorption experiment (top) and after the moisture sorption experiment(bottom). After the moisture sorption experiment, XRPD analysis revealedthat Form A is not stable and had converted to a new Form, Form K(Example 3). The x-axis is 2Theta measured in degrees and the y-axis isintensity measured in counts.

FIG. 5B is a comparison of XRPD patterns of Form D before the moisturesorption experiment (top) and after the moisture sorption experiment(bottom). After the moisture sorption experiment, XRPD analysis revealedthat Form D is not stable and had converted to a new Form, Form K(Example 3). The x-axis is 2Theta measured in degrees and the y-axis isintensity measured in counts.

FIG. 6 is a comparison of the XRPD patterns of Form A, Form B, and FormC after the stability study (Example 4) to reference Form A, Form B andForm C. The top thee patterns are reference forms of Form A, Form B, andForm C. After the seven-day stability study, Form A converted to a newForm (Form A post-study), but after equilibrium at room temperature for3 days, the new form changed back to Form A (Form A after 3 days). FormB and Form C remained unchanged during the stability study. The x-axisis 2Theta measured in degrees and the y-axis is intensity measured incounts.

FIG. 7 is the XRPD pattern for pure Form B. The peaks, marked with bars,are listed in Example 6. The x-axis is 2Theta measured in degrees andthe y-axis is intensity measured in counts.

FIG. 8 is a comparison of the XRPD patterns of impure Form B materialand pure Form B material as characterized in Example 6. Impure Form Bmaterial has two peaks at approximately 4.0 and 5.6 degrees that aremissing in the pure Form B material. The x-axis is 2Theta measured indegrees and the y-axis is relative intensity as a means to compare thetwo Form B materials.

FIG. 9 is a comparison of impure Form B, Samples 1 and 4 from the slurryexperiment described in Example 8, and pure Form B. Pure Form B is theForm B characterized in Example 6. A number of experiments wereconducted to convert impure Form B to pure Form B material, including aslurry experiment with 1:1 (v/v) 0.1 M HCl:acetone (Sample 1) and 1:2(v/v) 0.5 M HCl:acetone (Sample 4). The acidic aqueous acetone mixturesfailed to convert impure material to pure material. The XRPD patterns ofSample 1 and 4 were not consistent with the XRPD pattern of pure Form Bsince a peak at approximately 4.0 degrees was still present. The x-axisis relative humidity measured as a percent and the y-axis is weight ofwater of the material measured as a percent.

FIG. 10 is a comparison of impure samples of Form B, Samples 3 and 5from the slurry experiment described in Example 8, and pure Form B. Anumber of experiments were conducted to convert impure Form B to pureForm B material, including a slurry experiment with 75:25 (v/v) 0.1 MHCl:acetone (Sample 3) and 50:50 (v/v) 0.5 M HCl:acetone (Sample 5). Theacidic aqueous acetone mixtures failed to convert impure material topure material. Pure Form B is the Form B characterized in Example 6.Impure Form B is the material used as starting material in the slurryexperiments and impure Form B Sample 2 is a second impure form used as areference. The x-axis is relative humidity measured as a percent and they-axis is weight of water of the material measured as a percent.

FIG. 11 is a comparison of impure Form B, Samples 6, 7, and 11 from theslurry experiment described in Example 8, and pure Form B. A number ofexperiments were conducted to convert impure Form B to pure Form Bmaterial, including slurry experiments with 1:2 (v/v) water:acetone thatstirred at room temperature. Samples 6, 7, and 11 varied in theconcentration of impure Form B and the length of time that the samplesstirred (details are given in Table 12). All three conditions convertedimpure Form B to pure Form B since the XRPD patterns from Samples 6, 7,and 11 matched the pure Form B XRPD pattern. Pure Form B is the Form Bcharacterized in Example 6 and impure Form B is the material used asstaring material in the slurry experiments. The x-axis is relativehumidity measured as a percent and the y-axis is weight of water of thematerial measured as a percent.

FIG. 12 is a comparison of impure Form B, Samples 12 and 14 from theslurry experiment described in Example 8, and pure Form B. Slurryexperiments with 1:3 (v/v) water:acetone (Sample 14) and 1:2water:acetone followed by additional acetone (Sample 12) were conductedin an effort to improve the yield of the recrystallization process. TheXRPD patterns of Samples 12 and 14 were not consistent with the XRPDpattern of Form B since a peak at approximately 4.0 degrees was stillpresent. Pure Form B is the Form B characterized in Example 6 and impureForm B is the material used as staring material in the slurryexperiments. The x-axis is relative humidity measured as a percent andthe y-axis is weight of water of the material measured as a percent.

FIG. 13 is a graph from the TG-IR experiment of pure Form B, Sample 11(Example 8). The TG data showed a 6.4% wt loss at 33-137° C. The x-axisis temperature measured in degrees Celsius and the y-axis is weight ofthe material measured as a percent.

FIG. 14 is IR data from the TG-IR experiment of pure Form B, Sample 11(Example 8). The x-axes are wavenumber measured in cm⁻¹ and timemeasured in minutes. The y-axis is absorbance.

FIG. 15 compares IR spectra of pure Form B, Sample 11 obtained at 2.691minutes and 5.382 minutes in the TG-IR experiment to IR spectra of waterand hydrogen chloride. During the TG-IR experiment, only water, and nohydrogen chloride, was released as a volatile. The x-axis is wavenumbermeasured in cm⁻¹ and the y-axis is absorbance.

FIG. 16 is a comparison of Sample 8 dried in a vacuum oven for 15 hoursat approximately 40° C. (Example 8, Table 15). The XRPD following thevacuum procedure did not correlate with the XRPD pattern of pure Form B.Dry sample 8 is a new crystalline Form. The x-axis is 2Theta measured indegrees and the y-axis is intensity measured in counts.

FIG. 17 are the XRPD patterns of Sample 11 and Sample 23 that were bothdried in a vacuum oven, but under different conditions (Example 8, Table15) compared to the XRPD pattern of pure Form B. Both Sample 11 and 23exhibited XRPD patterns of Form B. The x-axis is 2Theta measured indegrees and the y-axis is intensity measured in counts.

FIG. 18 is a comparison of XRPD patterns from impure Form B, pure FormB, and the material that was converted from impure Form B as describedin Example 8. The XRPD pattern of the converted material aligned withthe pure Form B material. Pure Form B is the Form B characterized inExample 6 and impure Form B is the material used as staring material inthe conversion procedure. The x-axis is 2Theta measured in degrees andthe y-axis is intensity measured in counts.

FIG. 19 is the TGA data from the batch converted to pure Form B materialfrom impure Form B as described in Example 8. The TGA data showed a 7.6%weight loss at 31-120° C. it also showed an approximately 20% weightloss from 120-350° C. The x-axis is temperature measured in degreesCelsius and the y-axis is weight of the material measured as a percent.

FIG. 20 are XRPD patterns of Form I and Form J. The x-axis is 2Thetameasured in degrees and the y-axis is intensity measured in counts.

FIG. 21 is the DSC data from a representative batch of Form B material.The DSC data was collected by increasing the temperature of the sample(3.9 mg) from 25-400° C. at a rate of 10° C./minute. Endotherms wereobserved at 113° C. (1), 231° C. (2), 262° C. (3), and 348° C. (4).Endotherm 1 (integral=−237 mJ; normalized=−60 J/g) exhibited an onset of113° C. and an endset of 140° C. Endotherm 2 (integral=−182 mJ;normalized=−46 J/g) exhibited an onset of 219° C. and an endset of 239°C. Endotherm 3 (integral=177 mJ; normalized=45 J/g) exhibited an onsetof 250° C. and an endset of 271° C. Endotherm 4 (integral=−728 mJ;normalized=−186 J/g) exhibited an onset of 341° C. and an endset of 350°C. The x-axis is temperature measured in Celsius and the y-axis the heatflow measured in milli Watts (mW).

FIG. 22 is the DSC data from a representative batch of Form A. The DSCdata was collected by increasing the temperature of the sample (4.4 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at110° C. (1), 275° C. (2), and 344° C. (3). Endotherm 1 (integral=−670mJ; normalized=−151 J/g) exhibited an onset of 84° C. Endotherm 2(integral=−480 mJ; normalized=−108 J/g) exhibited an onset of 242° C.Endotherm 3 exhibited an onset of 344° C. The x-axis is temperaturemeasured in Celsius and the y-axis the heat flow measured in milli Watts(mW).

FIG. 23 is the DSC data from a representative batch of Form B. The DSCdata was collected by increasing the temperature of the sample (2.6 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at95° C. (1), 225° C. (2), 254° C. (3), and 348° C. (4). Endotherm 1(integral=−256 mJ; normalized=−97 J/g) exhibited an onset of 75° C.Endotherm 2 (integral=−265 mJ; normalized=−101 J/g) exhibited an onsetof 199° C. Endotherm 3 (integral=−140 mJ; normalized=−53 J/g) exhibitedan onset of 239° C. Endotherm 4 (integral=−94 mJ; normalized=−36 J/g)exhibited an onset of 344° C. The x-axis is temperature measured inCelsius and the y-axis the heat flow measured in milli Watts (mW).

FIG. 24 is the DSC data from a representative batch of Form C. The DSCdata was collected by increasing the temperature of the sample (2.5 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at95° C. (1), 235° C. (2), 257° C. (3), and 344° C. (4). Endotherm 1(integral=−88 mJ; normalized=−36 J/g) exhibited an onset of 77° C.Endotherm 2 (integral=−58 mJ; normalized=−23 J/g) exhibited an onset of216° C. Endotherm 3 (integral=−31 mJ; normalized=−12 J/g) exhibited anonset of 247° C. Endotherm 4 (integral=−379 mJ; normalized=−154 J/g)exhibited an onset of 338° C. The x-axis is temperature measured inCelsius and the y-axis the heat flow measured in milli Watts (mW).

FIG. 25 is the DSC data from a representative batch of Form D. The DSCdata was collected by increasing the temperature of the sample (2.5 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at103° C. (1), 260° C. (2) and 345° C. (3). Endotherm 1 (integral=−370 mJ;normalized=−149 J/g) exhibited an onset of 73° C. Endotherm 2(integral=−271 mJ; normalized=−109 J/g) exhibited an onset of 228° C.Endotherm 3 (integral=−321 mJ; normalized=−129 J/g) exhibited an onsetof 340° C. The x-axis is temperature measured in Celsius and the y-axisthe heat flow measured in milli Watts (mW).

FIG. 26 is the DSC data from a representative batch of Form E. The DSCdata was collected by increasing the temperature of the sample (2.5 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at70° C. (1), 219° C. (2), 275° C. (3), and 345° C. (4). Endotherm 1(integral=−495 mJ; normalized=−194 J/g) exhibited an onset of 38° C.Endotherm 2 (integral=25 mJ; normalized=10 J/g) exhibited an onset of209° C. Endotherm 3 (integral=−208 mJ; normalized=−81 J/g) exhibited anonset of 242° C. Endotherm 4 (integral=−339 mJ; normalized=−133 J/g)exhibited an onset of 340° C. The x-axis is temperature measured inCelsius and the y-axis the heat flow measured in milli Watts (mW).

FIG. 27 is the DSC data from a representative batch of Form F. The DSCdata was collected by increasing the temperature of the sample (3.0 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at73° C. (1), 214° C. (2), 277° C. (3), 303° C. (4), and 329° C. (5).Endotherm 1 (integral=−991 mJ; normalized=−323 J/g) exhibited an onsetof 43° C. Endotherm 2 (integral=−121 mJ; normalized=−39 J/g) exhibitedan onset of 205° C. Endotherm 3 (integral=98 mJ; normalized=32 J/g)exhibited an onset of 265° C. Endotherm 4 (integral=−15 mJ;normalized=−5 J/g) exhibited an onset of 297° C. Endotherm 5(integral=−283 mJ; normalized=−92 J/g) exhibited an onset of 318° C. Thex-axis is temperature measured in Celsius and the y-axis the heat flowmeasured in milli Watts (mW).

FIG. 28 is the DSC data from a representative batch of Form G. The DSCdata was collected by increasing the temperature of the sample (2.8 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at81° C. (1), 120° C. (2), 260° C. (3), and 347° C. (4). Endotherm 1(integral=−167 mJ; normalized=−59 J/g) exhibited an onset of 56° C.Endotherm 2 (integral=−183 mJ; normalized=−65 J/g) exhibited an onset of103° C. Endotherm 3 (integral=−251 mJ; normalized=−89 J/g) exhibited anonset of 235° C. Endotherm 4 (integral=−164 mJ; normalized=−58 J/g)exhibited an onset of 344° C. The x-axis is temperature measured inCelsius and the y-axis the heat flow measured in milli Watts (mW).

FIG. 29 is the DSC data from a representative batch of Form H. The DSCdata was collected by increasing the temperature of the sample (2.7 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at110° C. (1), 225° C. (2), 274° C. (3), and 346° C. (4). Endotherm 1(integral=−300 mJ; normalized=−110 J/g) exhibited an onset of 109° C.Endotherm 2 (integral=−41 mJ; normalized=−15 J/g) exhibited an onset of210° C. Endotherm 3 (integral=−138 mJ; normalized=−50 J/g) exhibited anonset of 242° C. Endotherm 4 (integral=−301 mJ; normalized=−110 J/g)exhibited an onset of 346° C. The x-axis is temperature measured inCelsius and the y-axis the heat flow measured in milli Watts (mW).

FIG. 30 is the DSC data from a representative batch of Form A. The DSCdata was collected by increasing the temperature of the sample (6.0 mg)from 30-350° C. at a rate of 10° C./minute. Endotherms were observed at121° C. (1), 242° C. (2), 290° C. (3), and 348° C. (4). Endotherm 1(integral=−541 mJ; normalized=−90 J/g) exhibited an onset of 93° C.Endotherm 2 (integral=133 mJ; normalized=22 J/g) exhibited an onset of233° C. Endotherm 3 (integral=−272 mJ; normalized=−45 J/g) exhibited anonset of 268° C. Endotherm 4 (integral=−1131 mJ; normalized=−198 J/g)exhibited an onset of 344° C. The x-axis is temperature measured inCelsius and the y-axis the heat flow measured in milli Watts (mW).

FIG. 31 is the XRPD pattern for Form I and Form J. The x-axis is 2Thetameasured in degrees and the y-axis is intensity measured in counts.

DETAILED DESCRIPTION OF THE INVENTION

It cannot be predicted in advance whether a compound exists in more thanone solid form or what the various properties of any solid form might beif one or more does exist, or whether the properties are advantageousfor a therapeutic dosage form. As one example, the drug ritonavir isactive in one polymorphic form and inactive in another form, and theinactive form is the more stable.

Solid forms of compounds can be characterized by analytical methods suchas X-ray powder diffraction pattern (XRDP), thermogravimetric analysis(TGA), TGA with IR off-gas analysis, Differential Scanning calorimetry(DSC), melting point, FT-Raman spectroscopy, Dynamic Vapor Sorption(DVS), polarized light microscopy (PLM) or other techniques known in theart.

Eleven forms of Compound 2 were discovered from slurry andcrystallization experiments. Of these eleven forms, Form A, Form B, andForm D were found to have properties suitable for further development.Moisture sorption experiments revealed that Form B is an unexpectedsuperior crystalline stable solid.

Morphic Form B

Isolated morphic Form B of Compound 2 is provided in this invention.

In one embodiment, Form B is characterized by an XRPD pattern in orsubstantially similar to that set forth in FIG. 7 . In one embodiment,Form B is characterized by an XRPD pattern comprising at least three2theta values selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°,18.1±0.2°, 19.7±0.2°, and 22.4±0.2°. In one embodiment, Form B ischaracterized by an XRPD pattern comprising a peak with a 2theta valueof 9.5±0.4°.

In one embodiment, Form B is characterized as having a 7.5% weight lossbetween 31 and 120° C. in a thermogravimetric infrared (TG-IR) analysis.

In one embodiment the isolated Compound 2 Form B does not have a peak atone or at both of 4.0±0.2° and 5.6±0.2° 2Theta, or the peak at one or atboth of 4.0±0.2° and 5.6±0.2° 2Theta is not greater than 200, 150, 100,or 75 Counts Per Second (CPS).

Form B can be prepared using selective crystallization. The method canbe carried out by treating a solution comprising a suitable solvent(s)and Compound 2 optionally in the presence of one or more seedscomprising Form B to conditions that provide for the crystallization ofForm B. The selective crystallization can be carried out in any suitablesolvent. For example, it can be carried out in an aprotic solvent or amixture thereof. The selective crystallization can be carried out at,for example, a temperature in the range of about 40° C. to about 65° C.In another embodiment the selective crystallization can be carried outat, for example, a temperature in the range of about 45° C. to about 60°C. or about 45° C. to about 55° C.

In one embodiment, Compound 2 Form B is produced by recrystallization ina solution of hydrochloric acid. Compound 1 is dissolved in aqueous HCland heated to at least 55±10° C. The solution is stirred forapproximately 45 minutes and filtered through an in-line filter. Acetoneis slowly added to the hot solution to induce crystallization. Thetemperature of the solution is then decreased to 25±5° C. or lower andstirred for at least 2 hours. The resulting solids are collected viafiltration to afford Form B.

In an alternative embodiment, Compound 2 Form B is recrystallized fromCompound 2 Form D. Compound 2 Form D is first formed by dissolvingCompound 1 in aqueous HCl and heating the solution to about 55±10° C.The solution is stirred for approximately 45 minutes and the resultingsolution is filtered through an in-line filter. The temperature of thesolution is then decreased to about 25±5° C. and the solution is stirredfor at least 2 hours. Acetone is added at a temperature of about 25±5°C. over the course of about one hour after crystallization has begun todrive crystallization to completion. The solution is stirred for aboutan additional 2 hours and the resulting solids are collected viafiltration to afford Compound 2 Form D. Form D is then dissolved inconcentrated HCl and the solution is heated. Acetone is added to the hotsolution prior to the formation of any solids. As the solution cools,the solids are collected via filtration to afford Form B.

In one embodiment, impure Compound 2 Form B is converted to pure Form Bin a water:acetone (1:2) (v/v) slurry at 30° C. This is followed by slowfiltration that results in a wet cake. The wet cake is dried at ambientconditions for about 3.5 hours followed by vacuum drying at ambienttemperature.

In certain embodiments, Form B is characterized by an XRPD patterncomprising all or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, or 20 2theta values selected from:

-   -   a. 6.5, 8.1, 9.4, 9.6, 10.2, 10.6, 11.2, 12.2, 12.9, 13.0, 13.3,        13.4, 14.0, 14.4, 14.6, 15.0, 15.9, 16.2, 16.4, 16.5, 16.8,        18.1, 18.4, 18.5, 18.6, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4,        19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 20.6, 21.3, 21.4, 21.8,        22.0, 22.2, 22.3, 22.4, 22.5, 22.8, 23.0, 23.1, 23.4, 23.8,        24.1, 24.2, 24.3, 24.4, 24.5, 24.6, 25.4, 25.6, 25.7, 25.9,        26.0, 26.1, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.2,        27.3, 27.5, 27.6, 27.7, 27.9, 28.3, 28.4, 28.5, 28.7, 28.9,        29.0, 29.1, 29.3, 29.4, 29.5, 29.6, 29.7, 29.8, 29.9, 30.0,        30.3, 30.4, 30.5, 30.6, 30.7, 30.9, 31.2, 31.5, 31.6, 31.7,        31.8, 31.9, 32.0, 32.2, 32.3, 32.4, 32.5, 32.6, 32.7, 32.8,        33.1, 33.2, 33.3, 33.6, 33.7, 33.8, 34.0, 34.1, 34.2, 34.3,        34.6, 34.7, 34.8, 35.0 35.2, 35.3, 35.5, 35.6, 35.9, 36.0, 36.2,        36.5, 36.6, 36.7, 36.8, 36.9, 37.1, 37.2, 37.3, 37.4, 37.5,        37.6, 37.7, 37.8, 37.9, 38.2, 38.3, 38.4, 38.5, 38.6, 38.7,        38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5, 39.6, 39.7,        39.8, 39.9, and 40.0° 2θ; or    -   b. 6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 13.3, 13.4, 14.0, 14.4, 14.6,        15.0, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.9,        19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4,        22.3, 22.4, 22.5, 22.8, 23.0, 23.1, 23.4, 23.8, 26.3, 26.4,        26.5, 26.6, 26.7, 26.8, 26.9, 27.2, 27.3, 27.5, 27.6, 27.7,        27.9, 28.3, 28.4, 28.5, 28.7, 28.9, 29.0, 29.1, 29.3, 29.4,        29.5, 29.6, 29.7, 29.8, 29.9, and 30.0, ° 2θ; or    -   c. 6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 13.3, 13.4, 14.0, 14.4, 14.6,        15.0, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.9,        19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4,        22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8,        26.9, 27.7, 27.9, 27.9, 29.0, 29.1, 29.3, 29.4, 29.5, 29.6,        29.7, 29.8, 29.9, and 30.0, ° 2θ; or    -   d. 6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 13.3, 13.4, 14.0, 14.4, 14.6,        15.0, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.9,        19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4,        22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8,        26.9, 27.7, 27.9, and 27.9, ° 2θ; or    -   e. 6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 14.0, 14.4, 14.6, 15.0, 16.2,        16.4, 16.5, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3,        19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5,        22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and        27.9, ° 2θ; or    -   f. 6.5, 9.5, 14.0, 14.4, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9,        19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4,        22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8,        26.9, 27.7, 27.9, and 27.9, ° 2θ; or    -   g. 9.5, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3,        19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5,        22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and        27.9, ° 2θ; or    -   h. 9.5, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.2, 19.3, 19.4,        19.5, 19.6, 19.7, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4,        26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, ° 2θ; or    -   i. 9.5, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.2, 19.3, 19.4,        19.5, 19.6, 19.7, 22.3, 22.4, 22.5, 26.3, 26.4, 26.5, 26.6,        26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, ° 2θ; or    -   j. 9.5, 18.1, 18.4, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 22.3,        22.4, 22.5, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7,        27.9, and 27.9, ° 2θ; or    -   k. 9.5, 18.1, 18.4, 19.3, 19.7, 22.3, 22.4, 22.5, 26.3, 26.4,        26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, ° 2θ; or    -   l. 9.5, 18.1, 18.4, 19.3, 19.7, 22.4, 26.3, 26.4, 26.5, 26.6,        26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, ° 2θ; or    -   m. 9.5, 18.1, 18.4, 19.3, 19.7, 22.4, 26.6, 27.7, 27.9, and        27.9, ° 2θ; or    -   n. 9.5, 18.1, 18.4, 19.3, 19.7, 22.4, 26.6, and 27.7, ° 2θ; or    -   o. 9.5, 18.1, 19.3, 19.7, 22.4, 26.6, and 27.7, ° 2θ; or    -   p. 9.5, 18.1, 19.3, 22.4, 26.6, and 27.7, ° 2θ; or    -   q. any of the above peak lists wherein the ° 2θ are ±0.1; or    -   r. any of the above peak lists wherein the ° 2θ are ±0.2;    -   s. any of the above peak lists wherein the ° 2θ are ±0.3;    -   t. any of the above peak lists wherein the ° 2θ are ±0.4;    -   u. any of the above peak lists wherein the ° 2θ of the peak at        9.5 is +0.4;    -   v. any of the above peak lists wherein the ° 2θ of the peak at        9.5 is ±0.4 and the remaining peaks are ±0.1′20;    -   w. any of the above peak lists wherein the ° 2θ of the peak at        9.5 is +0.4 and the remaining peaks are ±0.2° 2θ;

In one embodiment Form B is characterized by an XRPD pattern describedabove and is further characterized by having no peaks of greater than200 CPS in between 4 and 6° 2θ. In one embodiment Form B ischaracterized by an XRPD pattern described above and is furthercharacterized by having no peaks of greater than 150 CPS in between 4and 6° 2θ. In one embodiment Form B is characterized by an XRPD patterndescribed above and is further characterized by having no peaks ofgreater than 100 CPS in between 4 and 6° 2θ. In one embodiment Form B ischaracterized by an XRPD pattern described above and is furthercharacterized by having no peaks of greater than 75 CPS in between 4 and6° 2θ.

In one embodiment Form B is characterized by an XRPD pattern describedabove and is further characterized by having no peak of about 4.0° 2θ ofgreater than 150 CPS. In one embodiment Form B is characterized by anXRPD pattern described above and is further characterized by having nopeak of about 4.0° 2θ of greater than 100 CPS. In one embodiment Form Bis characterized by an XRPD pattern described above and is furthercharacterized by having no peak of about 4.0° 2θ greater than 75 CPS.

In one embodiment Form B is characterized by an XRPD pattern describedabove and is further characterized by having no peak of about 5.6° 2θ ofgreater than 150 CPS. In one embodiment Form B is characterized by anXRPD pattern described above and is further characterized by having nopeak of about 5.6° 2θ of greater than 100 CPS. In one embodiment Form Bis characterized by an XRPD pattern described above and is furthercharacterized by having no peak of about 5.6° 2θ greater than 75 CPS inbetween 4 and 6° 2θ.

In one embodiment Form B is characterized by an XRPD pattern describedabove and is further characterized by having no peak of about 5.3° 2θ ofgreater than 150 CPS. In one embodiment Form B is characterized by anXRPD pattern described above and is further characterized by having nopeak of about 5.3° 2θ of greater than 100 CPS. In one embodiment Form Bis characterized by an XRPD pattern described above and is furthercharacterized by having no peak of about 5.3° 2θ greater than 75 CPS inbetween 4 and 6° 2θ.

In a further embodiment, the CPS counts above are base-line corrected.

Methods utilized in preparing Form B are further described in Example 2and Example 8 below.

Morphic Form D

In one embodiment, Form D is characterized by DSC onset endotherms atabout 100±20° C., about 270±20° C., and about 347±20° C., for example at108.3° C., 266.1° C., and 347.0° C. or 95° C., 257° C., and 344° C.

Form D can be prepared using selective crystallization. The method canbe carried out by treating a solution comprising a suitable solvent(s)and Compound 2 optionally in the presence of one or more seedscomprising Form D to conditions that provide for the crystallization ofForm D. The selective crystallization can be carried out in any suitablesolvent. For example, it can be carried out in an aprotic solvent or amixture thereof. In one embodiment, the solvent is acetonitrile. Theselective crystallization can be carried out at, for example, atemperature in the range of about 5° C. to about 55° C.

In one embodiment, Compound 2 Form D is formed by dissolving Compound 1in aqueous 2M HCl (10 volumes) and heating the solution to 55±10° C. Thesolution is stirred for approximately 45 minutes and the resultingsolution is filtered through an in-line filter. The temperature of thesolution is then decreased to 25±5° C. and the solution is stirred forat least 2 hours. Acetone (30 volumes) is added at a temperature of25±5° C. over the course of an hour after crystallization has begun todrive crystallization to completion. The solution is stirred for anadditional 2 hours and the resulting solids are collected via filtrationto afford Compound 2 Form D.

In an alternative embodiment, Compound 2 Form D is formed by dissolvingCompound 1 in aqueous 2M HCl (10 volumes) and heating the solution to55±10° C. The solution is stirred for 45 minutes and the resultingsolution is filtered through an in-line filter. The solution is cooledto 25±5° C. and the solution is stirred for at least 2 hours. Theresulting solids are collected via filtration and acetone is added toafford Compound 2 Form D.

In one embodiment, Form D is again recrystallized to produce Form B.

Methods utilized in preparing Form D are further described in Example 2below.

Morphic Form A

In one embodiment, Forma A is characterized by an XRPD peaks at about of7.4±0.2 and 9.0±0.2 2theta. In an additional embodiment, Form A ischaracterized by DSC onset endotherms at about 110±20° C., about 275±20°C., and about 350±20° C., for example at 110.3° C., 275.6° C., and344.8° C. or 103° C., 260° C., and 345° C.

Form A can be prepared using selective crystallization. The method canbe carried out by treating a solution comprising a suitable solvent(s)and Compound 2 optionally in the presence of one or more seedscomprising Form A to conditions that provide for the crystallization ofForm D. The selective crystallization can be carried out in any suitablesolvent. For example, it can be carried out in a protic solvent or amixture thereof. In one embodiment, the solvent is MeOH, EtOH, or1-BuOH. The selective crystallization can be carried out at, forexample, a temperature in the range of about 5° C. to about 75° C. Inone embodiment, the crystallization is carried out at a temperature ofabout 60° C.

Methods utilized in preparing Form A are further described in Example 2below.

Chemical Description and Terminology

Compounds are described using standard nomenclature. Unless definedotherwise, all technical and scientific terms used herein have the samemeaning as is commonly understood by one of skill in the art to whichthis invention belongs.

The terms “a” and “an” do not denote a limitation of quantity, butrather denote the presence of at least one of the referenced item. Theterm “or” means “and/or”. Recitation of ranges of values are merelyintended to serve as a shorthand method of referring individually toeach separate value falling within the range, unless otherwise indicatedherein, and each separate value is incorporated into the specificationas if it were individually recited herein. The endpoints of all rangesare included within the range and independently combinable.

All methods described herein can be performed in any suitable orderunless otherwise indicated herein or otherwise clearly contradicted bycontext. The use of any and all examples, or exemplary language (e.g.,“such as”), is intended merely for illustration and does not pose alimitation on the scope of the invention unless otherwise claimed.

An “active agent” is a compound (including a compound disclosed herein),element, or mixture that when administered to a patient, alone or incombination with another compound, element, or mixture, confers,directly or indirectly, a physiological effect on the patient. Theindirect physiological effect may occur via a metabolite or otherindirect mechanism.

“Deuteration” and “deuterated” means that a hydrogen is replaced by adeuterium such that the deuterium exists over natural abundance and isthus “enriched”. An enrichment of 50% means that rather than hydrogen atthe specified position the deuterium content is 50%. For clarity, it isconfirmed that the term “enriched” as used herein does not meanpercentage enriched over natural abundance. In other embodiments, therewill be at least 80%, at least 90%, or at least 95% deuterium enrichmentat the specified deuterated position or positions. In other embodimentsthere will be at least 96%, at least 97%, at least 98%, or at least 99%deuterium enrichment at the specified deuterated position or positionsindicated. In the absence of indication to the contrary, the enrichmentof deuterium in the specified position of the compound described hereinis at least 90%.

A “dosage form” means a unit of administration of an active agent.Non-limiting examples of dosage forms include tablets, capsules,injections, suspensions, liquids, intravenous fluids, emulsions, creams,ointments, suppositories, inhalable forms, transdermal forms, and thelike.

“Pharmaceutical compositions” are compositions comprising at least oneactive agent, such as a compound or salt of one of the active compoundsdisclosed herein, and at least one other substance, such as a carrier.Pharmaceutical compositions optionally contain more than one activeagent. “Pharmaceutical combinations” or “combination therapy” refers tothe administration of at least two active agents, and in one embodiment,three or four or more active agents which may be combined in a singledosage form or provided together in separate dosage forms optionallywith instructions that the active agents are to be used together totreat a disorder.

“Pharmaceutically acceptable salts” includes derivatives of thedisclosed compounds in which the parent compound is modified by makinginorganic and organic, suitably non-toxic, acid or base addition saltsthereof. The salts of the present compounds can be synthesized from aparent compound that contains a basic or acidic moiety by conventionalchemical methods. Generally, such salts can be prepared by reacting freeacid forms of these compounds with a stoichiometric amount of theappropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate,bicarbonate, or the like), or by reacting free base forms of thesecompounds with a stoichiometric amount of the appropriate acid. Suchreactions are typically carried out in water or in an organic solvent,or in a mixture of the two. The pharmaceutically acceptable salt can bein the form of a pure crystal, or single morphic form, or can be used innon-crystalline or amorphic, glassy, or vitreous form, or a mixturethereof. In an alternative embodiment, the active compound can beprovided in the form of a solvate.

Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts and the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, conventional non-toxic acid salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,HOOC—(CH2)n-COOH where n is 0-4, and the like. Lists of additionalsuitable salts may be found, e.g., in Remington's PharmaceuticalSciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418(1985).

The term “carrier” means a diluent, excipient, or vehicle with which anactive compound is provided.

A “pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition/combination that isgenerally safe, is sufficiently non-toxic, and neither biologically norotherwise undesirable. A “pharmaceutically acceptable excipient” as usedin the present application includes both one and more than one suchexcipient.

A “patient” or “host” is a human or non-human animal, including, but notlimited to, simian, avian, feline, canine, bovine, equine or porcine inneed of medical treatment. Medical treatment can include treatment of anexisting condition, such as a disease or disorder, or a prophylactic ordiagnostic treatment. In a particular embodiment, the patient or host isa human patient. In an alternative embodiment, the patient such as ahost is treated to prevent a disorder or disease described herein.

The term “isolated” as used herein refers to the material insubstantially pure form. An isolated compound does not have anothercomponent that materially affects the properties of the compound. Inparticular embodiments, an isolated form is at least 50, 60, 70, 80, 90,95, 98 or 99% pure.

Methods of Treatment

In one aspect, a method of treating a proliferative disorder in a host,including a human, is provided comprising administering isolatedCompound 2 morphic Form B as described herein optionally in apharmaceutically acceptable carrier. Non-limiting examples of disordersinclude tumors, cancers, disorders related to abnormal cellularproliferation, inflammatory disorders, immune disorders, and autoimmunedisorders.

Compound 2 morphic Form B is useful as a therapeutic agent in a dosageform when administered in an effective amount to a host, including ahuman, to treat a tumor, cancer (solid, non-solid, diffuse,hematological, etc.), abnormal cellular proliferation, immune disorder,inflammatory disorder, blood disorder, a myelo- or lymphoproliferativedisorder such as B- or T-cell lymphomas, multiple myeloma, breastcancer, prostate cancer, AML, ALL, ACL, lung cancer, pancreatic cancer,colon cancer, skin cancer, melanoma, Waldenstrom's macroglobulinemia,Wiskott-Aldrich syndrome, or a post-transplant lymphoproliferativedisorder; an autoimmune disorder, for example, Lupus, Crohn's Disease,Addison disease, Celiac disease, dermatomyositis, Graves disease,thyroiditis, multiple sclerosis, pernicious anemia, reactive arthritis,or type I diabetes; a disease of cardiologic malfunction, includinghypercholesterolemia; an infectious disease, including a viral and/orbacterial infection; an inflammatory condition, including asthma,chronic peptic ulcers, tuberculosis, rheumatoid arthritis,periodontitis, ulcerative colitis, or hepatitis.

Exemplary proliferative disorders include, but are not limited to,benign growths, neoplasms, tumors, cancer (Rb positive or Rb negative),autoimmune disorders, inflammatory disorders graft-versus-hostrejection, and fibrotic disorders.

Non-limiting examples of cancers that can be treated according to thepresent invention include, but are not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangi osarcoma, lymphangioendotheliosarcoma, hemangiosarcoma),appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g.,cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast), brain cancer (e.g., meningioma;glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchuscancer, carcinoid tumor, cervical cancer (e.g., cervicaladenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma,colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma(e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma),endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophagealcancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),Ewing's sarcoma, eye cancer (e.g., intraocular melanoma,retinoblastoma), familiar hypereosinophilia, gall bladder cancer,gastric cancer (e.g., stomach adenocarcinoma), gastrointestinal stromaltumor (GIST), head and neck cancer (e.g., head and neck squamous cellcarcinoma, oral cancer (e.g., oral squamous cell carcinoma (OSCC),throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngealcancer, oropharyngeal cancer)), hematopoietic cancers (e.g., leukemiasuch as acute lymphocytic leukemia (ALL)—also known as acutelymphoblastic leukemia or acute lymphoid leukemia (e.g., B-cell ALL,T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cellAML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML),and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL);lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) andnon-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large celllymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)),follicular lymphoma, chronic lymphocytic leukemia/small lymphocyticlymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-celllymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas,nodal marginal zone B-cell lymphoma, splenic marginal zone B-celllymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma,lymphoplasmacytic lymphoma (i.e., “Waldenstrom's macroglobulinemia”),hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursorB-lymphoblastic lymphoma and primary central nervous system (CNS)lymphoma; and T-cell NHL such as precursor T-lymphoblasticlymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneousT-cell lymphoma (CTCL) (e.g., mycosis fungiodes, Sezary syndrome),angioimmunoblastic T-cell lymphoma, extranodal natural killer T-celllymphoma, enteropathy type T-cell lymphoma, subcutaneouspanniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); amixture of one or more leukemia/lymphoma as described above; andmultiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease,gamma chain disease, mu chain disease), hemangioblastoma, inflammatorymyofibroblastic tumors, immunocytic amyloidosis, kidney cancer (e.g.,nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma), liver cancer(e.g., hepatocellular cancer (HCC), malignant hepatoma), lung cancer(e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-smallcell lung cancer (NSCLC), adenocarcinoma of the lung), leiomyosarcoma(LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplasticsyndrome (MDS), mesothelioma, myeloproliferative disorder (MPD) (e.g.,polycythemia Vera (PV), essential thrombocytosis (ET), agnogenic myeloidmetaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathicmyelofibrosis, chronic myelocytic leukemia (CIVIL), chronic neutrophilicleukemia (CNL), hypereosinophilic syndrome (HES)), neuroblastoma,neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2,schwannomatosis), neuroendocrine cancer (e.g., gastroenteropancreaticneuroendoctrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovariancancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarianadenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g.,pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm(IPMN), Islet cell tumors), penile cancer (e.g., Paget's disease of thepenis and scrotum), pinealoma, primitive neuroectodermal tumor (PNT),prostate cancer (e.g., prostate adenocarcinoma), rectal cancer,rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamouscell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cellcarcinoma (BCC)), small bowel cancer (e.g., appendix cancer), softtissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma,malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma,fibrosarcoma, myxosarcoma), sebaceous gland carcinoma, sweat glandcarcinoma, synovioma, testicular cancer (e.g., seminoma, testicularembryonal carcinoma), thyroid cancer (e.g., papillary carcinoma of thethyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer),urethral cancer, vaginal cancer and vulvar cancer (e.g., Paget's diseaseof the vulva).

In another embodiment, the disorder is myelodysplastic syndrome (MDS).

In certain embodiments, the cancer is a hematopoietic cancer. In certainembodiments, the hematopoietic cancer is a lymphoma. In certainembodiments, the hematopoietic cancer is a leukemia. In certainembodiments, the leukemia is acute myelocytic leukemia (AML).

In certain embodiments, the proliferative disorder is amyeloproliferative neoplasm. In certain embodiments, themyeloproliferative neoplasm (MPN) is primary myelofibrosis (PMF).

In certain embodiments, the cancer is a solid tumor. A solid tumor, asused herein, refers to an abnormal mass of tissue that usually does notcontain cysts or liquid areas. Different types of solid tumors are namedfor the type of cells that form them. Examples of classes of solidtumors include, but are not limited to, sarcomas, carcinomas, andlymphomas, as described above herein. Additional examples of solidtumors include, but are not limited to, squamous cell carcinoma, coloncancer, breast cancer, prostate cancer, lung cancer, liver cancer,pancreatic cancer, and melanoma.

In certain embodiments, the condition treated with Compound 2 morphicForm B is a disorder related to abnormal cellular proliferation.

Abnormal cellular proliferation, notably hyperproliferation, can occuras a result of a wide variety of factors, including genetic mutation,infection, exposure to toxins, autoimmune disorders, and benign ormalignant tumor induction.

There are a number of skin disorders associated with cellularhyperproliferation. Psoriasis, for example, is a benign disease of humanskin generally characterized by plaques covered by thickened scales. Thedisease is caused by increased proliferation of epidermal cells ofunknown cause. Chronic eczema is also associated with significanthyperproliferation of the epidermis. Other diseases caused byhyperproliferation of skin cells include atopic dermatitis, lichenplanus, warts, pemphigus vulgaris, actinic keratosis, basal cellcarcinoma and squamous cell carcinoma.

Other hyperproliferative cell disorders include blood vesselproliferation disorders, fibrotic disorders, autoimmune disorders,graft-versus-host rejection, tumors and cancers.

Blood vessel proliferative disorders include angiogenic and vasculogenicdisorders. Proliferation of smooth muscle cells in the course ofdevelopment of plaques in vascular tissue cause, for example,restenosis, retinopathies and atherosclerosis. Both cell migration andcell proliferation play a role in the formation of atheroscleroticlesions.

Fibrotic disorders are often due to the abnormal formation of anextracellular matrix. Examples of fibrotic disorders include hepaticcirrhosis and mesangial proliferative cell disorders. Hepatic cirrhosisis characterized by the increase in extracellular matrix constituentsresulting in the formation of a hepatic scar. Hepatic cirrhosis cancause diseases such as cirrhosis of the liver. An increasedextracellular matrix resulting in a hepatic scar can also be caused byviral infection such as hepatitis. Lipocytes appear to play a major rolein hepatic cirrhosis.

Mesangial disorders are brought about by abnormal proliferation ofmesangial cells. Mesangial hyperproliferative cell disorders includevarious human renal diseases, such as glomerulonephritis, diabeticnephropathy, malignant nephrosclerosis, thrombotic micro-angiopathysyndromes, transplant rejection, and glomerulopathies.

Another disease with a proliferative component is rheumatoid arthritis.Rheumatoid arthritis is generally considered an autoimmune disease thatis thought to be associated with activity of autoreactive T cells, andto be caused by autoantibodies produced against collagen and IgE.

Other disorders that can include an abnormal cellular proliferativecomponent include Bechet's syndrome, acute respiratory distress syndrome(ARDS), ischemic heart disease, post-dialysis syndrome, leukemia,acquired immune deficiency syndrome, vasculitis, lipid histiocytosis,septic shock and inflammation in general.

In certain embodiments a compound of the present invention and itspharmaceutically acceptable derivatives or pharmaceutically acceptableformulations containing these compounds are also useful in theprevention and treatment of HBV infections and other related conditionssuch as anti-HBV antibody positive and HBV-positive conditions, chronicliver inflammation caused by HBV, cirrhosis, acute hepatitis, fulminanthepatitis, chronic persistent hepatitis, and fatigue. These compounds orformulations can also be used prophylactically to prevent or retard theprogression of clinical illness in individuals who are anti-HBV antibodyor HBV-antigen positive or who have been exposed to HBV.

In certain embodiments, the condition is associated with an immuneresponse.

Cutaneous contact hypersensitivity and asthma are just two examples ofimmune responses that can be associated with significant morbidity.Others include atopic dermatitis, eczema, Sjogren's Syndrome, includingkeratoconjunctivitis sicca secondary to Sjogren's Syndrome, alopeciaareata, allergic responses due to arthropod bite reactions, Crohn'sdisease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis,ulcerative colitis, cutaneous lupus erythematosus, scleroderma,vaginitis, proctitis, and drug eruptions. These conditions may result inany one or more of the following symptoms or signs: itching, swelling,redness, blisters, crusting, ulceration, pain, scaling, cracking, hairloss, scarring, or oozing of fluid involving the skin, eye, or mucosalmembranes.

In atopic dermatitis, and eczema in general, immunologically mediatedleukocyte infiltration (particularly infiltration of mononuclear cells,lymphocytes, neutrophils, and eosinophils) into the skin importantlycontributes to the pathogenesis of these diseases. Chronic eczema alsois associated with significant hyperproliferation of the epidermis.Immunologically mediated leukocyte infiltration also occurs at sitesother than the skin, such as in the airways in asthma and in the tearproducing gland of the eye in keratoconjunctivitis sicca.

In one non-limiting embodiment compounds of the present invention areused as topical agents in treating contact dermatitis, atopicdermatitis, eczematous dermatitis, psoriasis, Sjogren's Syndrome,including keratoconjunctivitis sicca secondary to Sjogren's Syndrome,alopecia areata, allergic responses due to arthropod bite reactions,Crohn's disease, aphthous ulcer, iritis, conjunctivitis,keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma,cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, anddrug eruptions. The novel method may also be useful in reducing theinfiltration of skin by malignant leukocytes in diseases such as mycosisfungoides. These compounds can also be used to treat anaqueous-deficient dry eye state (such as immune mediatedkeratoconjunctivitis) in a patient suffering therefrom, by administeringthe compound topically to the eye.

The term “neoplasia” or “cancer” is used throughout the specification torefer to the pathological process that results in the formation andgrowth of a cancerous or malignant neoplasm, i.e., abnormal tissue(solid) or cells (non-solid) that grow by cellular proliferation, oftenmore rapidly than normal and continues to grow after the stimuli thatinitiated the new growth cease. Malignant neoplasms show partial orcomplete lack of structural organization and functional coordinationwith the normal tissue and most invade surrounding tissues, canmetastasize to several sites, are likely to recur after attemptedremoval and may cause the death of the patient unless adequatelytreated. As used herein, the term neoplasia is used to describe allcancerous disease states and embraces or encompasses the pathologicalprocess associated with malignant hematogenous, ascitic and solidtumors. Exemplary cancers which may be treated by the present disclosedcompounds either alone or in combination with at least one additionalanticancer agent include squamous-cell carcinoma, basal cell carcinoma,adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas,cancer of the bladder, bowel, breast, cervix, colon, esophagus, head,kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach;leukemias; benign and malignant lymphomas, particularly Burkitt'slymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas;myeloproliferative diseases; sarcomas, including Ewing's sarcoma,hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheralneuroepithelioma, synovial sarcoma, gliomas, astrocytomas,oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas,ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors,meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowelcancer, breast cancer, prostate cancer, cervical cancer, uterine cancer,lung cancer, ovarian cancer, testicular cancer, thyroid cancer,astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, livercancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease,Wilms' tumor and teratocarcinomas. Additional cancers which may betreated using the disclosed compounds according to the present inventioninclude, for example, acute granulocytic leukemia, acute lymphocyticleukemia (ALL), acute myelogenous leukemia (AML), adenocarcinoma,adenosarcoma, adrenal cancer, adrenocortical carcinoma, anal cancer,anaplastic astrocytoma, angiosarcoma, appendix cancer, astrocytoma,Basal cell carcinoma, B-Cell lymphoma, bile duct cancer, bladder cancer,bone cancer, bone marrow cancer, bowel cancer, brain cancer, brain stemglioma, breast cancer, triple (estrogen, progesterone and HER-2)negative breast cancer, double negative breast cancer (two of estrogen,progesterone and HER-2 are negative), single negative (one of estrogen,progesterone and HER-2 is negative), estrogen-receptor positive,HER2-negative breast cancer, estrogen receptor-negative breast cancer,estrogen receptor positive breast cancer, metastatic breast cancer,luminal A breast cancer, luminal B breast cancer, Her2-negative breastcancer, HER2-positive or negative breast cancer, progesteronereceptor-negative breast cancer, progesterone receptor-positive breastcancer, recurrent breast cancer, carcinoid tumors, cervical cancer,cholangiocarcinoma, chondrosarcoma, chronic lymphocytic leukemia (CLL),chronic myelogenous leukemia (CIVIL), colon cancer, colorectal cancer,craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuseastrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer,ependymoma, epithelioid sarcoma, esophageal cancer, ewing sarcoma,extrahepatic bile duct cancer, eye cancer, fallopian tube cancer,fibrosarcoma, gallbladder cancer, gastric cancer, gastrointestinalcancer, gastrointestinal carcinoid cancer, gastrointestinal stromaltumors (GIST), germ cell tumor glioblastoma multiforme (GBM), glioma,hairy cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkinlymphoma, hypopharyngeal cancer, infiltrating ductal carcinoma (IDC),infiltrating lobular carcinoma (ILC), inflammatory breast cancer (IBC),intestinal Cancer, intrahepatic bile duct cancer, invasive/infiltratingbreast cancer, Islet cell cancer, jaw cancer, Kaposi sarcoma, kidneycancer, laryngeal cancer, leiomyosarcoma, leptomeningeal metastases,leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma insitu, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma,male breast cancer, medullary carcinoma, medulloblastoma, melanoma,meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma,mesenchymous, mesothelioma metastatic breast cancer, metastatic melanomametastatic squamous neck cancer, mixed gliomas, monodermal teratoma,mouth cancer mucinous carcinoma, mucosal melanoma, multiple myeloma,Mycosis Fungoides, myelodysplastic syndrome, nasal cavity cancer,nasopharyngeal cancer, neck cancer, neuroblastoma, neuroendocrine tumors(NETs), non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oatcell cancer, ocular cancer, ocular melanoma, oligodendroglioma, oralcancer, oral cavity cancer, oropharyngeal cancer, osteogenic sarcoma,osteosarcoma, ovarian cancer, ovarian epithelial cancer ovarian germcell tumor, ovarian primary peritoneal carcinoma, ovarian sex cordstromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma,paranasal sinus cancer, parathyroid cancer, pelvic cancer, penilecancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer,pheochromocytoma, pilocytic astrocytoma, pineal region tumor,pineoblastoma, pituitary gland cancer, primary central nervous system(CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma,renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, softtissue sarcoma, bone sarcoma, sarcoma, sinus cancer, skin cancer, smallcell lung cancer (SCLC), small intestine cancer, spinal cancer, spinalcolumn cancer, spinal cord cancer, squamous cell carcinoma, stomachcancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throatcancer, thymoma/thymic carcinoma, thyroid cancer, tongue cancer, tonsilcancer, transitional cell cancer, tubal cancer, tubular carcinoma,undiagnosed cancer, ureteral cancer, urethral cancer, uterineadenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvarcancer, T-cell lineage acute lymphoblastic leukemia (T-ALL), T-celllineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, AdultT-cell leukemia, Pre-B ALL, Pre-B lymphomas, large B-cell lymphoma,Burkitts lymphoma, B-cell ALL, Philadelphia chromosome positive ALL,Philadelphia chromosome positive CML, juvenile myelomonocytic leukemia(JMML), acute promyelocytic leukemia (a subtype of AML), large granularlymphocytic leukemia, Adult T-cell chronic leukemia, diffuse large Bcell lymphoma, follicular lymphoma; Mucosa-Associated Lymphatic Tissuelymphoma (MALT), small cell lymphocytic lymphoma, mediastinal large Bcell lymphoma, nodal marginal zone B cell lymphoma (NMZL); splenicmarginal zone lymphoma (SMZL); intravascular large B-cell lymphoma;primary effusion lymphoma; or lymphomatoid granulomatosis; B-cellprolymphocytic leukemia; splenic lymphoma/leukemia, unclassifiable,splenic diffuse red pulp small B-cell lymphoma; lymphoplasmacyticlymphoma; heavy chain diseases, for example, Alpha heavy chain disease,Gamma heavy chain disease, Mu heavy chain disease, plasma cell myeloma,solitary plasmacytoma of bone; extraosseous plasmacytoma; primarycutaneous follicle center lymphoma, T cell/histocyte rich large B-celllymphoma, DLBCL associated with chronic inflammation; Epstein-Barr virus(EBV)+ DLBCL of the elderly; primary mediastinal (thymic) large B-celllymphoma, primary cutaneous DLBCL, leg type, ALK+ large B-cell lymphoma,plasmablastic lymphoma; large B-cell lymphoma arising in HHV8-associatedmulticentric, Castleman disease; B-cell lymphoma, unclassifiable, withfeatures intermediate between diffuse large B-cell lymphoma, or B-celllymphoma, unclassifiable, with features intermediate between diffuselarge B-cell lymphoma and classical Hodgkin lymphoma.

In another aspect, a method of increasing BIM expression (e.g., BCLC2L11expression) is provided to induce apoptosis in a cell comprisingcontacting a compound of the present invention or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof withthe cell. In certain embodiments, the method is an in vitro method. Incertain embodiments, the method is an in vivo method. BCL2L11 expressionis tightly regulated in a cell. BCL2L11 encodes for BIM, a proapoptoticprotein. BCL2L11 is downregulated in many cancers and BIM is inhibitedin many cancers, including chronic myelocytic leukemia (CML) andnon-small cell lung cancer (NSCLC) and that suppression of BCL2L11expression can confer resistance to tyrosine kinase inhibitors. See,e.g., Ng et al., Nat. Med. (2012) 18:521-528.

In yet another aspect, a method of treating a condition associated withangiogenesis is provided, such as, for example, a diabetic condition(e.g., diabetic retinopathy), an inflammatory condition (e.g.,rheumatoid arthritis), macular degeneration, obesity, atherosclerosis,or a proliferative disorder, comprising administering to a subject inneed thereof a compound of the present invention or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof.

In certain embodiments, the condition associated with angiogenesis ismacular degeneration. In certain embodiments, provided is a method oftreating macular degeneration comprising administering to a subject inneed thereof a compound of the present invention or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof.

In certain embodiments, the condition associated with angiogenesis isobesity. As used herein, “obesity” and “obese” as used herein, refers toclass I obesity, class II obesity, class III obesity and pre-obesity(e.g., being “over-weight”) as defined by the World Health Organization.In certain embodiments, a method of treating obesity is providedcomprising administering to a subject in need thereof a compound of thepresent invention or a pharmaceutically acceptable composition, salt,isotopic analog, or prodrug thereof.

In certain embodiments, the condition associated with angiogenesis isatherosclerosis. In certain embodiments, provided is a method oftreating atherosclerosis comprising administering to a subject in needthereof a compound of the present invention or a pharmaceuticallyacceptable composition, salt, isotopic analog, or prodrug thereof.

In certain embodiments, the condition associated with angiogenesis is aproliferative disorder. In certain embodiments, provided is a method oftreating a proliferative disorder comprising administering to a subjectin need thereof a compound of the present invention or apharmaceutically acceptable composition, salt, isotopic analog, orprodrug thereof.

In an alternative embodiment Compound 2 Form A or D is administered inan effective amount to treat a proliferative disorder.

In another alternative embodiment Compound 2 Form C, E, G, or H isadministered in an effective amount to treat a proliferative disorder.

Methods to Reduce the Side Effects Related to Chemotherapy

In certain embodiments, the isolated Compound 2 Form B of the presentinvention decreases the effect of chemotherapeutic agent toxicity onCDK4/6 replication dependent healthy cells, such as hematopoietic stemcells and hematopoietic progenitor cells (together referred to asHSPCs), and/or renal epithelial cells, in subjects, typically humans,that will be, are being, or have been exposed to the chemotherapeuticagent (typically a DNA-damaging agent).

In one embodiment, the subject has been exposed to a chemotherapeuticagent, and, using the isolated Compound 2 Form B described herein, thesubject's CDK4/6-replication dependent healthy cells are placed in G1arrest following exposure in order to mitigate, for example, DNA damage.In one embodiment, the compound is administered at least ½ hour, atleast 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, atleast 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, atleast 10 hours, at least 12 hours, at least 14 hours, at least 16 hours,at least 18 hours, at least 20 hours or more post chemotherapeutic agentexposure.

In one embodiment, the isolated Compound 2 Form B can allow for doseintensification (e.g., more therapy can be given in a fixed period oftime) in medically related chemotherapies, which will translate tobetter efficacy. Therefore, the presently disclosed methods can resultin chemotherapy regimens that are less toxic and more effective.

In some embodiments, the use of the isolated Compound 2 Form B describedherein may result in reduced or substantially free of off-targeteffects, for example, related to inhibition of kinases other than CDK4and/or CDK6 such as CDK2. Furthermore, in certain embodiments, the useof the isolated Compound 2 Form B described herein should not inducecell cycle arrest in CDK4/6 replication independent cells.

In some embodiments, the use of the isolated Compound 2 Form B describedherein reduces the risk of undesirable off-target effects including, butnot limited to, long term toxicity, anti-oxidant effects, and estrogeniceffects. Anti-oxidant effects can be determined by standard assays knownin the art. For example, a compound with no significant anti-oxidanteffects is a compound that does not significantly scavengefree-radicals, such as oxygen radicals. The anti-oxidant effects of acompound can be compared to a compound with known anti-oxidant activity,such as genistein. Thus, a compound with no significant anti-oxidantactivity can be one that has less than about 2, 3, 5, 10, 30, or 100fold anti-oxidant activity relative to genistein. Estrogenic activitiescan also be determined via known assays. For instance, a non-estrogeniccompound is one that does not significantly bind and activate theestrogen receptor. A compound that is substantially free of estrogeniceffects can be one that has less than about 2, 3, 5, 10, 20, or 100 foldestrogenic activity relative to a compound with estrogenic activity,e.g., genistein.

In an alternative embodiment Compound 2 Form A or D is administered inan effective amount to decrease the effect of chemotherapeutic agenttoxicity on CDK4/6 replication dependent healthy cells, such ashematopoietic stem cells and hematopoietic progenitor cells (togetherreferred to as HSPCs), and/or renal epithelial cells, in subjects,typically humans, that will be, are being, or have been exposed to thechemotherapeutic agent (typically a DNA-damaging agent).

In an alternative embodiment Compound 2 Form C, E, G, or H isadministered in an effective amount to decrease the effect ofchemotherapeutic agent toxicity on CDK4/6 replication dependent healthycells, such as hematopoietic stem cells and hematopoietic progenitorcells (together referred to as HSPCs), and/or renal epithelial cells, insubjects, typically humans, that will be, are being, or have beenexposed to the chemotherapeutic agent (typically a DNA-damaging agent).

Methods to Treat Abnormal Proliferation of T-Cells, B-Cells, and/orNK-Cells

In certain aspects, the invention includes the use of an effectiveamount of the isolated Compound 2 Form B, or its pharmaceuticallyacceptable salt, prodrug or isotopic variant optionally in apharmaceutical composition, to treat a host, typically a human, with aselected cancer, tumor, hyperproliferative condition or an inflammatoryor immune disorder. Compound 2 Form B is also active against T-cellproliferation. Given the paucity of drugs for T-cell cancers andabnormal proliferation, the identification of such uses represents asubstantial improvement in the medical therapy for these diseases.

Abnormal proliferation of T-cells, B-cells, and/or NK-cells can resultin a wide range of diseases such as cancer, proliferative disorders andinflammatory/immune diseases. A host, for example a human, afflictedwith any of these disorders can be treated with an effective amount ofthe isolated Compound 2 Form B as described herein to achieve a decreasein symptoms (a palliative agent) or a decrease in the underlying disease(a disease modifying agent).

Examples include T-cell or NK-cell lymphoma, for example, but notlimited to: peripheral T-cell lymphoma; anaplastic large cell lymphoma,for example anaplastic lymphoma kinase (ALK) positive, ALK negativeanaplastic large cell lymphoma, or primary cutaneous anaplastic largecell lymphoma; angioimmunoblastic lymphoma; cutaneous T-cell lymphoma,for example mycosis fungoides, Sézary syndrome, primary cutaneousanaplastic large cell lymphoma, primary cutaneous CD30+ T-celllymphoproliferative disorder; primary cutaneous aggressiveepidermotropic CD8+ cytotoxic T-cell lymphoma; primary cutaneousgamma-delta T-cell lymphoma; primary cutaneous small/medium CD4+ T-celllymphoma, and lymphomatoid papulosis; Adult T-cell Leukemia/Lymphoma(ATLL); Blastic NK-cell Lymphoma; Enteropathy-type T-cell lymphoma;Hematosplenic gamma-delta T-cell Lymphoma; Lymphoblastic Lymphoma; NasalNK/T-cell Lymphomas; Treatment-related T-cell lymphomas; for examplelymphomas that appear after solid organ or bone marrow transplantation;T-cell prolymphocytic leukemia; T-cell large granular lymphocyticleukemia; Chronic lymphoproliferative disorder of NK-cells; AggressiveNK cell leukemia; Systemic EBV+ T-cell lymphoproliferative disease ofchildhood (associated with chronic active EBV infection); Hydroavacciniforme-like lymphoma; Adult T-cell leukemia/lymphoma;Enteropathy-associated T-cell lymphoma; Hepatosplenic T-cell lymphoma;or Subcutaneous panniculitis-like T-cell lymphoma.

In one embodiment, the isolated Compound 2 Form B as disclosed herein,or its salt, prodrug, or isotopic variant can be used in an effectiveamount to treat a host, for example a human, with a lymphoma orlymphocytic or myelocytic proliferation disorder or abnormality. Forexample, the isolated Compound 2 Form B as described herein can beadministered to a host suffering from a Hodgkin Lymphoma or aNon-Hodgkin Lymphoma. For example, the host can be suffering from aNon-Hodgkin Lymphoma such as, but not limited to: an AIDS-RelatedLymphoma; Anaplastic Large-Cell Lymphoma; Angioimmunoblastic Lymphoma;Blastic NK-Cell Lymphoma; Burkitt's Lymphoma; Burkitt-like Lymphoma(Small Non-Cleaved Cell Lymphoma); Chronic Lymphocytic Leukemia/SmallLymphocytic Lymphoma; Cutaneous T-Cell Lymphoma; Diffuse Large B-CellLymphoma; Enteropathy-Type T-Cell Lymphoma; Follicular Lymphoma;Hepatosplenic Gamma-Delta T-Cell Lymphoma; Lymphoblastic Lymphoma;Mantle Cell Lymphoma; Marginal Zone Lymphoma; Nasal T-Cell Lymphoma;Pediatric Lymphoma; Peripheral T-Cell Lymphomas; Primary Central NervousSystem Lymphoma; T-Cell Leukemias; Transformed Lymphomas;Treatment-Related T-Cell Lymphomas; or Waldenstrom's Macroglobulinemia.

Alternatively, the isolated Compound 2 Form B disclosed herein, or itssalt, prodrug, or isotopic variant can be used in an effective amount totreat a host, for example a human, with a Hodgkin Lymphoma, such as, butnot limited to: Nodular Sclerosis Classical Hodgkin's Lymphoma (CHL);Mixed Cellularity CHL; Lymphocyte-depletion CHL; Lymphocyte-rich CHL;Lymphocyte Predominant Hodgkin Lymphoma; or Nodular LymphocytePredominant HL.

Alternatively, the isolated Compound 2 Form B disclosed herein, or itssalt, prodrug, or isotopic variant can be used in an effective amount totreat a host, for example a human with a specific B-cell lymphoma orproliferative disorder such as, but not limited to: multiple myeloma;Diffuse large B cell lymphoma; Follicular lymphoma; Mucosa-AssociatedLymphatic Tissue lymphoma (MALT); Small cell lymphocytic lymphoma;Mediastinal large B cell lymphoma; Nodal marginal zone B cell lymphoma(NMZL); Splenic marginal zone lymphoma (SMZL); Intravascular largeB-cell lymphoma; Primary effusion lymphoma; or Lymphomatoidgranulomatosis; B-cell prolymphocytic leukemia; Hairy cell leukemia;Splenic lymphoma/leukemia, unclassifiable; Splenic diffuse red pulpsmall B-cell lymphoma; Hairy cell leukemia-variant; Lymphoplasmacyticlymphoma; Heavy chain diseases, for example, Alpha heavy chain disease,Gamma heavy chain disease, Mu heavy chain disease; Plasma cell myeloma;Solitary plasmacytoma of bone; Extraosseous plasmacytoma; Primarycutaneous follicle center lymphoma; T cell/histiocyte rich large B-celllymphoma; DLBCL associated with chronic inflammation; Epstein-Barr virus(EBV)+ DLBCL of the elderly; Primary mediastinal (thymic) large B-celllymphoma; Primary cutaneous DLBCL, leg type; ALK+ large B-cell lymphoma;Plasmablastic lymphoma; Large B-cell lymphoma arising in HHV8-associatedmulticentric; Castleman disease; B-cell lymphoma, unclassifiable, withfeatures intermediate between diffuse large B-cell lymphoma; or B-celllymphoma, unclassifiable, with features intermediate between diffuselarge B-cell lymphoma and classical Hodgkin lymphoma.

In one embodiment, the isolated Compound 2 Form B disclosed herein, orits salt, prodrug, or isotopic variant can be used in an effectiveamount to treat a host, for example a human with leukemia. For example,the host may be suffering from an acute or chronic leukemia of alymphocytic or myelogenous origin, such as, but not limited to: Acutelymphoblastic leukemia (ALL); Acute myelogenous leukemia (AML); Chroniclymphocytic leukemia (CLL); Chronic myelogenous leukemia (CML); juvenilemyelomonocytic leukemia (JMML); hairy cell leukemia (HCL); acutepromyelocytic leukemia (a subtype of AML); large granular lymphocyticleukemia; or Adult T-cell chronic leukemia. In one embodiment, thepatient suffers from an acute myelogenous leukemia, for example anundifferentiated AML (M0); myeloblastic leukemia (M1; with/withoutminimal cell maturation); myeloblastic leukemia (M2; with cellmaturation); promyelocytic leukemia (M3 or M3 variant [M3V]);myelomonocytic leukemia (M4 or M4 variant with eosinophilia [M4E]);monocytic leukemia (M5); erythroleukemia (M6); or megakaryoblasticleukemia (M7).

In an alternative embodiment Compound 2 Form A or D is administered inan effective amount to treat a host, typically a human, with a selectedcancer, tumor, hyperproliferative condition or an inflammatory or immunedisorder. Given the paucity of drugs for T-cell cancers and abnormalproliferation, the identification of such uses represents a substantialimprovement in the medical therapy for these diseases.

In an alternative embodiment Compound 2 Form C, E, G, or H isadministered in an effective amount to treat a host, typically a human,with a selected cancer, tumor, hyperproliferative condition or aninflammatory or immune disorder. Given the paucity of drugs for T-cellcancers and abnormal proliferation, the identification of such usesrepresents a substantial improvement in the medical therapy for thesediseases.

Pharmaceutical Compositions and Dosage Forms

The isolated Compound 2 Form B described herein, or an alternative salt,isotopic analog, or prodrug can be administered in an effective amountto a host to treat any of the disorders described herein using anysuitable approach which achieves the desired therapeutic result. Theamount and timing of the isolated Compound 2 Form B administered will,of course, be dependent on the host being treated, the instructions ofthe supervising medical specialist, on the time course of the exposure,on the manner of administration, on the pharmacokinetic properties ofthe particular active compound, and on the judgment of the prescribingphysician. Thus, because of host to host variability, the dosages givenbelow are a guideline and the physician can titrate doses of thecompound to achieve the treatment that the physician considersappropriate for the host. In considering the degree of treatmentdesired, the physician can balance a variety of factors such as age andweight of the host, presence of preexisting disease, as well as presenceof other diseases.

The pharmaceutical composition may be formulated as any pharmaceuticallyuseful form, e.g., a pill, a capsule, a tablet, a transdermal patch, asubcutaneous patch, a dry powder, an inhalation formulation, in amedical device, suppository, buccal, or sublingual formulation. Somedosage forms, such as tablets and capsules, are subdivided into suitablysized unit doses containing appropriate quantities of the activecomponents, e.g., an effective amount to achieve the desired purpose.

The therapeutically effective dosage of the isolated Compound 2 Form Bdescribed herein will be determined by the health care practitionerdepending on the condition, size and age of the patient as well as theroute of delivery. In one non-limited embodiment, a dosage from about0.1 to about 200 mg/kg has therapeutic efficacy, with all weights beingcalculated based upon the weight of the active compound. In someembodiments, the dosage may be the amount of the isolated Compound 2Form B needed to provide a serum concentration of the active compound ofup to about 10 nM, 50 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600nM, 700 nM, 800 nM, 900 nM, 1 μM, 5 μM, 10 μM, 20 μM, 30 μM, or 40 μM.

In certain embodiments the pharmaceutical composition is in a dosageform that contains from about 0.1 mg to about 2000 mg, from about 10 mgto about 1000 mg, from about 100 mg to about 800 mg, or from about 200mg to about 600 mg of the active compound and optionally from about 0.1mg to about 2000 mg, from about 10 mg to about 1000 mg, from about 100mg to about 800 mg, or from about 200 mg to about 600 mg of the isolatedCompound 2 Form B, measured alternatively either as the active compoundor its salt, in a unit dosage form. Examples of dosage forms with atleast 5, 10, 15, 20, 25, 50, 100, 200, 250, 300, 400, 500, 600, 700, or750 mg of active compound, or its salt. The pharmaceutical compositionmay also include a molar ratio of the isolated Compound 2 Form B and anadditional active agent, in a ratio that achieves the desired results.

The isolated Compound 2 Form B disclosed herein or used as describedherein may be administered orally, topically, parenterally, byinhalation or spray, sublingually, via implant, including ocularimplant, transdermally, via buccal administration, rectally,intramuscular, inhalation, intra-aortal, intracranial, subdermal,intraperitioneal, subcutaneous, transnasal, sublingual, or rectal or byother means, in dosage unit formulations containing conventionalpharmaceutically acceptable carriers.

In accordance with the presently disclosed methods, an oraladministration can be in any desired form in which the isolated Compound2 Form B is stable as a solid. In certain embodiments, the isolatedCompound 2 Form B is delivered in a solid microparticle or nanoparticle.When administered through inhalation the isolated Compound 2 Form B maybe in the form of a plurality of solid particles or droplets having anydesired particle size, and for example, from about 0.01, 0.1 or 0.5 toabout 5, 10, 20 or more microns, and optionally from about 1 to about 2microns. The isolated Compound 2 Form B as disclosed in the presentinvention has good pharmacokinetic and pharmacodynamics properties, forinstance when administered by the oral or intravenous routes.

The pharmaceutical formulations can comprise the isolated Compound 2Form B described herein or an alternative pharmaceutically acceptablesalt thereof, in any pharmaceutically acceptable carrier.

Carriers include excipients and diluents and must be of sufficientlyhigh purity and sufficiently low toxicity to render them suitable foradministration to the patient being treated. The carrier can be inert orit can possess pharmaceutical benefits of its own. The amount of carrieremployed in conjunction with the compound is sufficient to provide apractical quantity of material for administration per unit dose of thecompound.

Classes of carriers include, but are not limited to binders, bufferingagents, coloring agents, diluents, disintegrants, emulsifiers,flavorants, glidents, lubricants, preservatives, stabilizers,surfactants, tableting agents, and wetting agents. Some carriers may belisted in more than one class, for example vegetable oil may be used asa lubricant in some formulations and a diluent in others. Exemplarypharmaceutically acceptable carriers include sugars, starches,celluloses, powdered tragacanth, malt, gelatin; talc, and vegetableoils. Optional active agents may be included in a pharmaceuticalcomposition, which do not substantially interfere with the activity ofthe compound of the present invention.

Depending on the intended mode of administration, the pharmaceuticalcompositions can be in the form of solid form or a semi-solid dosageform that the isolated Compound 2 Form B is stable in, such as, forexample, tablets, suppositories, pills, capsules, powders, or the like,preferably in unit dosage form suitable for single administration of aprecise dosage. The compositions will include an effective amount of theselected drug in combination with a pharmaceutically acceptable carrierand, in addition, can include other pharmaceutical agents, adjuvants,diluents, buffers, and the like.

Thus, the compositions of the disclosure can be administered aspharmaceutical formulations including those suitable for oral (includingbuccal and sub-lingual), rectal, nasal, topical, pulmonary, vaginaladministration or in a form suitable for administration by inhalation orinsufflation. The preferred manner of administration is oral using aconvenient daily dosage regimen which can be adjusted according to thedegree of affliction. For solid compositions, conventional nontoxicsolid carriers include, for example, pharmaceutical grades of mannitol,lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose,glucose, sucrose, magnesium carbonate, and the like.

In yet another embodiment is the use of permeation enhancer excipientsincluding polymers such as: polycations (chitosan and its quaternaryammonium derivatives, poly-L-arginine, aminated gelatin); polyanions(N-carboxymethyl chitosan, poly-acrylic acid); and, thiolated polymers(carboxymethyl cellulose-cysteine, polycarbophil-cysteine,chitosan-thiobutylamidine, chitosan-thioglycolic acid,chitosan-glutathione conjugates).

For oral administration, the composition will generally take the form ofa tablet or capsule. Tablets and capsules are preferred oraladministration forms. Tablets and capsules for oral use can include oneor more commonly used carriers such as lactose and corn starch.Lubricating agents, such as magnesium stearate, are also typicallyadded. Typically, the compositions of the disclosure can be combinedwith an oral, non-toxic, pharmaceutically acceptable, inert carrier suchas lactose, starch, sucrose, glucose, methyl cellulose, magnesiumstearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol andthe like. Moreover, when desired or necessary, suitable binders,lubricants, disintegrating agents, and coloring agents can also beincorporated into the mixture. Suitable binders include starch, gelatin,natural sugars such as glucose or beta-lactose, corn sweeteners, naturaland synthetic gums such as acacia, tragacanth, or sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes, and the like.Lubricants used in these dosage forms include sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate, sodiumchloride, and the like. Disintegrators include, without limitation,starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

In addition to the active compounds or their salts, the pharmaceuticalformulations can contain other additives, such as pH-adjustingadditives. In particular, useful pH-adjusting agents include acids, suchas hydrochloric acid, bases or buffers, such as sodium lactate, sodiumacetate, sodium phosphate, sodium citrate, sodium borate, or sodiumgluconate. Further, the formulations can contain antimicrobialpreservatives. Useful antimicrobial preservatives include methylparaben,propylparaben, and benzyl alcohol. An antimicrobial preservative istypically employed when the formulations is placed in a vial designedfor multi-dose use. The pharmaceutical formulations described herein canbe lyophilized using techniques well known in the art.

For oral administration a pharmaceutical composition can take the formof a tablet, pill, capsule, powder, and the like. Tablets containingvarious excipients such as sodium citrate, calcium carbonate and calciumphosphate may be employed along with various disintegrants such asstarch (e.g., potato or tapioca starch) and certain complex silicates,together with binding agents such as polyvinylpyrrolidone, sucrose,gelatin and acacia. Additionally, lubricating agents such as magnesiumstearate, sodium lauryl sulfate, and talc are often very useful fortableting purposes. Solid compositions of a similar type may be employedas fillers in soft and hard-filled gelatin capsules.

Pharmaceutical formulations also are provided which provide a controlledrelease of a compound described herein, including through the use of adegradable polymer, as known in the art.

The term “pharmaceutically acceptable salts” as used herein refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with hosts (e.g., human hosts) without unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use,as well as the zwitterionic forms, where possible, of the compounds ofthe presently disclosed host matter.

In an alternative embodiment Compound 2 morphic form B is not a HClsalt, but is instead a salt described below.

In one embodiment the additional therapeutic agent described in theCombination Section below is administered as a pharmaceuticallyacceptable salt, for example, a salt described below.

Thus, the term “salts” refers to the relatively non-toxic, inorganic andorganic acid addition salts of the presently disclosed compounds. Thesesalts can be prepared during the final isolation and purification of thecompounds or by separately reacting the purified compound in its freebase form with a suitable organic or inorganic acid and isolating thesalt thus formed. Basic compounds are capable of forming a wide varietyof different salts with various inorganic and organic acids. Acidaddition salts of the basic compounds are prepared by contacting thefree base form with a sufficient amount of the desired acid to producethe salt in the conventional manner. The free base form can beregenerated by contacting the salt form with a base and isolating thefree base in the conventional manner. The free base forms may differfrom their respective salt forms in certain physical properties such assolubility in polar solvents. Pharmaceutically acceptable base additionsalts may be formed with metals or amines, such as alkali and alkalineearth metal hydroxides, or of organic amines. Examples of metals used ascations, include, but are not limited to, sodium, potassium, magnesium,calcium, and the like. Examples of suitable amines include, but are notlimited to, N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, N-methylglucamine, and procaine. Thebase addition salts of acidic compounds are prepared by contacting thefree acid form with a sufficient amount of the desired base to producethe salt in the conventional manner. The free acid form can beregenerated by contacting the salt form with an acid and isolating thefree acid in a conventional manner. The free acid forms may differ fromtheir respective salt forms somewhat in certain physical properties suchas solubility in polar solvents.

Salts can be prepared from inorganic acids sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, nitrate, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, chloride, bromide, iodide such as hydrochloric, nitric,phosphoric, sulfuric, hydrobromic, hydriodic, phosphorus, and the like.Representative salts include the hydrobromide, hydrochloride, sulfate,bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate,stearate, laurate, borate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate,glucoheptonate, lactobionate, laurylsulphonate and isethionate salts,and the like. Salts can also be prepared from organic acids, such asaliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoicacids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids,aliphatic and aromatic sulfonic acids, etc. and the like. Representativesalts include acetate, propionate, caprylate, isobutyrate, oxalate,malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate,benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate,benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate,maleate, tartrate, methanesulfonate, and the like. Pharmaceuticallyacceptable salts can include cations based on the alkali and alkalineearth metals, such as sodium, lithium, potassium, calcium, magnesium andthe like, as well as non-toxic ammonium, quaternary ammonium, and aminecations including, but not limited to, ammonium, tetramethylammonium,tetraethyl ammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. Also contemplated are the saltsof amino acids such as arginate, gluconate, galacturonate, and the like.See, for example, Berge et al., J. Pharm. Sci., 1977, 66, 1-19, which isincorporated herein by reference.

Formulations suitable for rectal administration are typically presentedas unit dose suppositories. These may be prepared by admixing the activedisclosed compound with one or more conventional solid carriers, forexample, cocoa butter, and then shaping the resulting mixture.

Formulations suitable for topical application to the skin preferablytake the form of an ointment, cream, lotion, paste, gel, spray, aerosol,or oil, which maintain the stability of the isolated Compound 2 Form B.Carriers which may be used include petroleum jelly, lanoline,polyethylene glycols, alcohols, transdermal enhancers, and combinationsof two or more thereof.

Formulations suitable for transdermal administration may be presented asdiscrete patches adapted to remain in intimate contact with theepidermis of the recipient for a prolonged period of time. Formulationssuitable for transdermal administration may also be delivered byiontophoresis (see, for example, Pharmaceutical Research 3 (6):318(1986)) and typically take the form of an optionally buffered aqueoussolution of the active compound. In one embodiment, microneedle patchesor devices are provided for delivery of drugs across or into biologicaltissue, particularly the skin. The microneedle patches or devices permitdrug delivery at clinically relevant rates across or into skin or othertissue barriers, with minimal or no damage, pain, or irritation to thetissue.

In an alternative embodiment, Compound 2 Form B is an HCl salt, forexample a mono-hydrochloride salt, an HCl salt with about 1hydrochloride units per Compound 2 unit, about 1.5 hydrochloride unitsper Compound 2 unit, or about 2 hydrochloride units per Compound 2 unit.

In one embodiment, “about” means±1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or10%. In one embodiment, Compound 2 Form B has about 2 HCl ions perCompound 2 molecule.

Formulations suitable for administration to the lungs can be deliveredby a wide range of passive breath driven and active power drivensingle/-multiple dose dry powder inhalers (DPI). The devices mostcommonly used for respiratory delivery include nebulizers, metered-doseinhalers, and dry powder inhalers. Several types of nebulizers areavailable, including jet nebulizers, ultrasonic nebulizers, andvibrating mesh nebulizers. Selection of a suitable lung delivery devicedepends on parameters, such as nature of the drug and its formulation,the site of action, and pathophysiology of the lung.

Combination Therapy

Isolated Compound 2 morphic Form B can be used in an effective amountalone or in combination with another compound of the present inventionor another bioactive agent to treat a host such as a human with adisorder as described herein.

The isolated Compound 2 Form B described herein can be used in aneffective amount alone or in combination with another compound of thepresent invention or another bioactive agent to treat a host such as ahuman with a disorder as described herein.

The term “bioactive agent” is used to describe an agent, other than theselected compound according to the present invention, which can be usedin combination or alternation with a compound of the present inventionto achieve a desired result of therapy. In one embodiment, the compoundof the present invention and the bioactive agent are administered in amanner that they are active in vivo during overlapping time periods, forexample, have time-period overlapping Cmax, Tmax, AUC or otherpharmacokinetic parameter. In another embodiment, isolated Compound 2Form B and the bioactive agent are administered to a host in needthereof that do not have overlapping pharmacokinetic parameter, however,one has a therapeutic impact on the therapeutic efficacy of the other.

In one aspect of this embodiment, the bioactive agent is an immunemodulator, including but not limited to a checkpoint inhibitor,including as non-limiting examples, a PD-1 inhibitor, PD-L1 inhibitor,PD-L2 inhibitor, CTLA-4 inhibitor, LAG-3 inhibitor, TIM-3 inhibitor,V-domain Ig suppressor of T-cell activation (VISTA) inhibitors, smallmolecule, peptide, nucleotide, or other inhibitor. In certain aspects,the immune modulator is an antibody, such as a monoclonal antibody.

PD-1 inhibitors that blocks the interaction of PD-1 and PD-L1 by bindingto the PD-1 receptor, and in turn inhibit immune suppression include,for example, nivolumab (Opdivo), pembrolizumab (Keytruda), pidilizumab,AMP-224 (AstraZeneca and Medlmmune), PF-06801591 (Pfizer), MEDI0680(AstraZeneca), PDR001 (Novartis), REGN2810 (Regeneron), SHR-12-1(Jiangsu Hengrui Medicine Company and Incyte Corporation), TSR-042(Tesaro), and the PD-L1/VISTA inhibitor CA-170 (Curis Inc.). PD-L1inhibitors that block the interaction of PD-1 and PD-L1 by binding tothe PD-L1 receptor, and in turn inhibits immune suppression, include forexample, atezolizumab (Tecentriq), durvalumab (AstraZeneca andMedImmune), KNO35 (Alphamab), and BMS-936559 (Bristol-Myers Squibb).CTLA-4 checkpoint inhibitors that bind to CTLA-4 and inhibits immunesuppression include, but are not limited to, ipilimumab, tremelimumab(AstraZeneca and MedImmune), AGEN1884 and AGEN2041 (Agenus). LAG-3checkpoint inhibitors, include, but are not limited to, BMS-986016(Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), IMP321 (PrimaBioMed), LAG525 (Novartis), and the dual PD-1 and LAG-3 inhibitor MGD013(MacroGenics). An example of a TIM-3 inhibitor is TSR-022 (Tesaro).

In yet another embodiment, isolated Compound 2 Form B as describedherein can be administered in an effective amount for the treatment ofabnormal tissue of the female reproductive system such as breast,ovarian, endometrial, or uterine cancer, in combination or alternationwith an effective amount of an estrogen inhibitor including but notlimited to a SERM (selective estrogen receptor modulator), a SERD(selective estrogen receptor degrader), a complete estrogen receptordegrader, or another form of partial or complete estrogen antagonist oragonist. Partial anti-estrogens like raloxifene and tamoxifen retainsome estrogen-like effects, including an estrogen-like stimulation ofuterine growth, and also, in some cases, an estrogen-like action duringbreast cancer progression which actually stimulates tumor growth. Incontrast, fulvestrant, a complete anti-estrogen, is free ofestrogen-like action on the uterus and is effective intamoxifen-resistant tumors. Non-limiting examples of anti-estrogencompounds are provided in WO 2014/19176 assigned to Astra Zeneca,WO2013/090921, WO 2014/203129, WO 2014/203132, and US2013/0178445assigned to Olema Pharmaceuticals, and U.S. Pat. Nos. 9,078,871,8,853,423, and 8,703, 810, as well as US 2015/0005286, WO 2014/205136,and WO 2014/205138. Additional non-limiting examples of anti-estrogencompounds include: SERMS such as anordrin, bazedoxifene, broparestriol,chlorotrianisene, clomiphene citrate, cyclofenil, lasofoxifene,ormeloxifene, raloxifene, tamoxifen, toremifene, and fulvestrant;aromatase inhibitors such as aminoglutethimide, testolactone,anastrozole, exemestane, fadrozole, formestane, and letrozole; andantigonadotropins such as leuprorelin, cetrorelix, allylestrenol,chloromadinone acetate, cyproterone acetate, delmadinone acetate,dydrogesterone, medroxyprogesterone acetate, megestrol acetate,nomegestrol acetate, norethisterone acetate, progesterone, andspironolactone. Other estrogenic ligands that can be used according tothe present invention are described in U.S. Pat. Nos. 4,418,068;5,478,847; 5,393,763; and 5,457,117, WO2011/156518, U.S. Pat. Nos.8,455,534 and 8,299,112, 9,078,871; 8,853,423; 8,703,810; US2015/0005286; and WO 2014/205138, US2016/0175289, US2015/0258080, WO2014/191726, WO 2012/084711; WO 2002/013802; WO 2002/004418; WO2002/003992; WO 2002/003991; WO 2002/003990; WO 2002/003989; WO2002/003988; WO 2002/003986; WO 2002/003977; WO 2002/003976; WO2002/003975; WO 2006/078834; U.S. Pat. No. 6,821,989; US 2002/0128276;U.S. Pat. No. 6,777,424; US 2002/0016340; U.S. Pat. Nos. 6,326,392;6,756,401; US 2002/0013327; U.S. Pat. Nos. 6,512,002; 6,632,834; US2001/0056099; U.S. Pat. Nos. 6,583,170; 6,479,535; WO 1999/024027; U.S.Pat. No. 6,005,102; EP 0802184; U.S. Pat. Nos. 5,998,402; 5,780,497,5,880,137, WO 2012/048058 and WO 2007/087684.

In another embodiment, the isolated Compound 2 Form B described hereincan be administered in an effective amount for the treatment of abnormaltissue of the male reproductive system such as prostate or testicularcancer, in combination or alternation with an effective amount of anandrogen (such as testosterone) inhibitor including but not limited to aselective androgen receptor modulator, a selective androgen receptordegrader, a complete androgen receptor degrader, or another form ofpartial or complete androgen antagonist. In one embodiment, the prostateor testicular cancer is androgen-resistant. Non-limiting examples ofanti-androgen compounds are provided in WO 2011/156518 and U.S. Pat.Nos. 8,455,534 and 8,299,112. Additional non-limiting examples ofanti-androgen compounds include: enzalutamide, apalutamide, cyproteroneacetate, chlormadinone acetate, spironolactone, canrenone, drospirenone,ketoconazole, topilutamide, abiraterone acetate, and cimetidine.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of abirateroneacetate (Zytiga) for the treatment of abnormal tissue of the malereproductive system.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of abirateroneacetate (Zytiga) for the treatment of prostate cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of enzalutamide forthe treatment of prostate cancer.

In one embodiment, the bioactive agent is an ALK inhibitor. Examples ofALK inhibitors include but are not limited to Crizotinib, Alectinib,ceritinib, TAE684 (NVP-TAE684), GSK1838705A, AZD3463, ASP3026,PF-06463922, entrectinib (RXDX-101), and AP26113. In one embodiment, thebioactive agent is an EGFR inhibitor. Examples of EGFR inhibitorsinclude erlotinib (Tarceva), gefitinib (Iressa), afatinib (Gilotrif),rociletinib (CO-1686), osimertinib (Tagrisso), olmutinib (Olita),naquotinib (ASP8273), nazartinib (EGF816), PF-06747775 (Pfizer),icotinib (BPI-2009), neratinib (HKI-272; PB272); avitinib (AC0010),EA1045, tarloxotinib (TH-4000; PR-610), PF-06459988 (Pfizer),tesevatinib (XL647; EXEL-7647; KD-019), transtinib, WZ-3146, WZ8040,CNX-2006, dacomitinib (PF-00299804; Pfizer), brigatinib (Alunbrig),lorlatinib, and PF-06747775 (PF7775).

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of afatinibdimaleate (Gilotrif) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of alectinib(Alecensa) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ceritinib(Zykadia) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of crizotinib(Xalkori) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of osimertinib(Tagrisso) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of brigatinib(Alunbrig) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of lorlatinib forthe treatment of non-small cell lung cancer.

In one embodiment, the bioactive agent is an HER-2 inhibitor. Examplesof HER-2 inhibitors include trastuzumab, lapatinib, ado-trastuzumabemtansine, and pertuzumab.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of lapatinibditosylate for the treatment of breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of lapatinibditosylate for the treatment of HER2+ breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of PF7775 for thetreatment of non-small cell lung cancer.

In one embodiment, the bioactive agent is a CD20 inhibitor. Examples ofCD20 inhibitors include obinutuzumab, rituximab, fatumumab, ibritumomab,tositumomab, and ocrelizumab.

In one embodiment, the bioactive agent is a JAK3 inhibitor. Examples ofJAK3 inhibitors include tasocitinib.

In one embodiment, the bioactive agent is a BCL-2 inhibitor. Examples ofBCL-2 inhibitors include venetoclax, ABT-199(4-[4-[[2-(4-Chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide),ABT-737(4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide) (navitoclax), ABT-263((R)-4-(4-((4′-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide),GX15-070 (obatoclax mesylate,(2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4-methoxypyrrol-2-ylidene]indole;methanesulfonic acid))), 2-methoxy-antimycin A3, YC137(4-(4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester),pogosin, ethyl2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate,Nilotinib-d3, TW-37(N-[4-[[2-(1,1-Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl]benzamide),Apogossypolone (ApoG2), HA14-1, AT101, sabutoclax, gambogic acid, orG3139 (Oblimersen).

In one aspect, a treatment regimen is provided comprising theadministration of Compound 2 morphic Form B in combination with at leastone additional chemotherapeutic agent. The combinations disclosed hereincan be administered for beneficial, additive, or synergistic effect inthe treatment of abnormal cellular proliferative disorders.

In specific embodiments, the treatment regimen includes theadministration of isolated Compound 2 morphic Form B in combination withat least one kinase inhibitor. In one embodiment, the at least onekinase inhibitor is selected from a phosphoinositide 3-kinase (PI3K)inhibitor, a Bruton's tyrosine kinase (BTK) inhibitor, or a spleentyrosine kinase (Syk) inhibitor, or a combination thereof.

PI3k inhibitors that may be used in the present invention are wellknown. Examples of PI3 kinase inhibitors include but are not limited toWortmannin, demethoxyviridin, perifosine, idelalisib, Pictilisib,Palomid 529, ZSTK474, PWT33597, CUDC-907, and AEZS-136, duvelisib,GS-9820, BKM120, GDC-0032 (Taselisib),(2-[4-[2-(2-Isopropyl-5-methyl-1,2,4-triazol-3-yl)-5,6-dihydroimidazo[1,2-d][1,4]benzoxazepin-9-yl]pyrazol-1-yl]-2-methylpropanamide),MLN-1117 ((2R)-1-Phenoxy-2-butanyl hydrogen (S)-methylphosphonate; orMethyl(oxo) {[(2R)-1-phenoxy-2-butanyl]oxy}phosphonium)), BYL-719((2S)-N1-[4-Methyl-5-[2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridinyl]-2-thiazolyl]-1,2-pyrrolidinedicarboxamide),GSK2126458 (2,4-Difluoro-N-{2-(methyloxy)-5-[4-(4-pyridazinyl)-6-quinolinyl]-3-pyridinyl}benzenesulfonamide)(omipalisib), TGX-221((±)-7-Methyl-2-(morpholin-4-yl)-9-(1-phenylaminoethyl)-pyrido[1,2-a]-pyrimidin-4-one),GSK2636771(2-Methyl-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylicacid dihydrochloride), KIN-193((R)-2-((1-(7-methyl-2-morpholino-4-oxo-4H-pyrido[1,2-a]pyrimidin-9-yl)ethyl)amino)benzoicacid), TGR-1202/RP5264, GS-9820((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-mohydroxypropan-1-one),GS-1101(5-fluoro-3-phenyl-2-([S)]-1-[9H-purin-6-ylamino]-propyl)-3H-quinazolin-4-one),AMG-319, GSK-2269557, SAR245409(N-(4-(N-(3-((3,5-dimethoxyphenyl)amino)quinoxalin-2-yl)sulfamoyl)phenyl)-3-methoxy-4methylbenzamide), BAY80-6946(2-amino-N-(7-methoxy-8-(3-morpholinopropoxy)-2,3-dihydroimidazo[1,2-c]quinaz),AS 252424(5-[1-[5-(4-Fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione),CZ 24832(5-(2-amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide),Buparlisib(5-[2,6-Di(4-morpholinyl)-4-pyrimidinyl]-4-(trifluoromethyl)-2-pyridinamine),GDC-0941(2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)-1-piperazinyl]methyl]-4-(4-morpholinyl)thieno[3,2-d]pyrimidine),GDC-0980((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (also known as RG7422)),SF1126((8S,14S,17S)-14-(carboxymethyl)-8-(3-guanidinopropyl)-17-(hydroxymethyl)-3,6,9,12,15-pentaoxo-1-(4-(4-oxo-8-phenyl-4H-chromen-2-yl)morpholino-4-ium)-2-oxa-7,10,13,16-tetraazaoctadecan-18-oate),PF-05212384(N-[4-[[4-(Dimethylamino)-1-piperidinyl]carbonyl]phenyl]-N′-[4-(4,6-di-4-morpholinyl-1,3,5-triazin-2-yl)phenyl]urea)(gedatolisib), LY3023414, BEZ235(2-Methyl-2-{4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl]phenyl}propanenitrile)(dactolisib), XL-765(N-(3-(N-(3-(3,5-dimethoxyphenylamino)quinoxalin-2-yl)sulfamoyl)phenyl)-3-methoxy-4-methylbenzamide),and GSK1059615(5-[[4-(4-Pyridinyl)-6-quinolinyl]methylene]-2,4-thiazolidenedione),PX886 ([(3aR,6E,9S,9aR,10R,11aS)-6-[[bis(prop-2-enyl)amino]methylidene]-5-hydroxy-9-(methoxymethyl)-9a,11a-dimethyl-1,4,7-trioxo-2,3,3a,9,10,11-hexahydroindeno[4,5h]isochromen-10-yl]acetate (also known as sonolisib)) LY294002, AZD8186, PF-4989216,pilaralisib, GNE-317, PI-3065, PI-103, NU7441 (KU-57788), HS 173,VS-5584 (SB2343), CZC24832, TG100-115, A66, YM201636, CAY10505, PIK-75,PIK-93, AS-605240, BGT226 (NVP-BGT226), AZD6482, voxtalisib, alpelisib,IC-87114, TGI100713, CH5132799, PKI-402, copanlisib (BAY 80-6946), XL147, PIK-90, PIK-293, PIK-294, 3-MA (3-methyladenine), AS-252424,AS-604850, apitolisib (GDC-0980; RG7422), and the structures describedin WO2014/071109. In one embodiment, isolated Compound 2 Form B iscombined in a single dosage form with the PIk3 inhibitor.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of alpelisib forthe treatment of solid tumors.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of alpelisib forthe treatment of abnormal tissue of the female reproductive system.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of alpelisib forthe treatment of breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of copanlisibhydrochloride (Aliqopa) for the treatment of lymphoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of copanlisibhydrochloride (Aliqopa) for the treatment of follicular lymphoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of idelalisib(Zydelig) for the treatment of chronic lymphocytic leukemia.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of idelalisib(Zydelig) for the treatment of Non-Hodgkin lymphoma, includingfollicular B-cell non-Hodgkin lymphoma or small lymphocytic lymphoma.

BTK inhibitors for use in the present invention are well known. Examplesof BTK inhibitors include ibrutinib (also known asPCI-32765)(Imbruvica™)(1-[(3R)-3-[4-amino-3-(4-phenoxy-phenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2-en-1-one),dianilinopyrimidine-based inhibitors such as AVL-101 and AVL-291/292(N-(3-((5-fluoro-2-((4-(2-methoxyethoxy)phenyl)amino)pyrimidin-4-yl)amino)phenyl)acrylamide) (Avila Therapeutics) (see US Patent Publication No 2011/0117073,incorporated herein in its entirety), Dasatinib([N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide],LFM-A13 (alpha-cyano-beta-hydroxy-beta-methyl-N-(2,5-ibromophenyl)propenamide), GDC-0834([R—N-(3-(6-(4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide],CGI-5604-(tert-butyl)-N-(3-(8-(phenylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide,CGI-1746(4-(tert-butyl)-N-(2-methyl-3-(4-methyl-6-((4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazin-2-yl)phenyl)benzamide),CNX-774(4-(4-(4-(3-acrylamidophenyl)amino)-5-fluoropyrimidin-2-yl)amino)phenoxy)-N-methylpicolinamide),CTA056(7-benzyl-1-(3-(piperidin-1-yl)propyl)-2-(4-(pyridin-4-yl)phenyl)-1H-imidazo[4,5-g]quinoxalin-6(5H)-one),GDC-0834((R)-N-(3-(6-((4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide),GDC-0837((R)-N-(3-(6-((4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide),HM-71224, ACP-196, ONO-4059 (Ono Pharmaceuticals), PRT062607(4-((3-(2H-1,2,3-triazol-2-yl)phenyl)amino)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamidehydrochloride), QL-47(1-(1-acryloylindolin-6-yl)-9-(1-methyl-1H-pyrazol-4-yl)benzo[h][1,6]naphthyridin-2(1H)-one),and RN486(6-cyclopropyl-8-fluoro-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridin-2-ylamino]-6-oxo-1,6-dihydro-pyridin-3-yl}-phenyl)-2H-isoquinolin-1-one),and other molecules capable of inhibiting BTK activity, for examplethose BTK inhibitors disclosed in Akinleye et ah, Journal of Hematology& Oncology, 2013, 6:59, the entirety of which is incorporated herein byreference. In one embodiment, an effective amount of the isolatedCompound 2 Form B is combined in a single dosage form with the BTKinhibitor.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ibrutinib(Imbruvica) for the treatment of chronic lymphocytic leukemia.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ibrutinib(Imbruvica) for the treatment of lymphoma, including small lymphocyticlymphoma, mantle cell lymphoma, marginal zone lymphoma, or Waldenströmmacroglobulinemia.

Syk inhibitors for use in the present invention are well known, andinclude, for example, Cerdulatinib(4-(cyclopropylamino)-2-((4-(4-(ethylsulfonyl)piperazin-1-yl)phenyl)amino)pyrimidine-5-carboxamide),entospletinib(6-(1H-indazol-6-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine),fostamatinib([6-({5-Fluoro-2-[(3,4,5-trimethoxyphenyl)amino]-4-pyrimidinyl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b][1,4]oxazin-4-yl]methyldihydrogen phosphate), fostamatinib disodium salt (sodium(64(5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-3-oxo-2H-pyrido[3,2-b][1,4]oxazin-4(3H)-yl)methylphosphate), BAY 61-3606(2-(7-(3,4-Dimethoxyphenyl)-imidazo[1,2-c]pyrimidin-5-ylamino)-nicotinamideHCl), R09021(6-[(1R,2S)-2-Amino-cyclohexylamino]-4-(5,6-dimethyl-pyridin-2-ylamino)-pyridazine-3-carboxylicacid amide), imatinib (Gleevac;4-[(4-methylpiperazin-1-yl)methyl]-N-(4-methyl-3-{[4-(pyridin-3-yl)pyrimidin-2-yl]amino}phenyl)benzamide),staurosporine, GSK143(2-(((3R,4R)-3-aminotetrahydro-2H-pyran-4-yl)amino)-4-(p-tolylamino)pyrimidine-5-carboxamide),PP2(1-(tert-butyl)-3-(4-chlorophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine),PRT-060318(2-(((1R,2S)-2-aminocyclohexyl)amino)-4-(m-tolylamino)pyrimidine-5-carboxamide),PRT-062607(4-((3-(2H-1,2,3-triazol-2-yl)phenyl)amino)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamidehydrochloride), R112(3,3′4(5-fluoropyrimidine-2,4-diyl)bis(azanediyl))diphenol), R348(3-Ethyl-4-methylpyridine), R406(6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one), piceatannol (3-Hydroxyresveratol), YM193306 (see Singh et al.Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J.Med. Chem. 2012, 55, 3614-3643), 7-azaindole, piceatannol, ER-27319 (seeSingh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK)Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 incorporated in itsentirety herein), Compound D (see Singh et al. Discovery and Developmentof Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55,3614-3643 incorporated in its entirety herein), PRT060318 (see Singh etal. Discovery and Development of Spleen Tyrosine Kinase (SYK)Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 incorporated in itsentirety herein), luteolin (see Singh et al. Discovery and Developmentof Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55,3614-3643 incorporated in its entirety herein), apigenin (see Singh etal. Discovery and Development of Spleen Tyrosine Kinase (SYK)Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 incorporated in itsentirety herein), quercetin (see Singh et al. Discovery and Developmentof Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55,3614-3643 incorporated in its entirety herein), fisetin (see Singh etal. Discovery and Development of Spleen Tyrosine Kinase (SYK)Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 incorporated in itsentirety herein), myricetin (see Singh et al. Discovery and Developmentof Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55,3614-3643 incorporated in its entirety herein), morin (see Singh et al.Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J.Med. Chem. 2012, 55, 3614-3643 incorporated in its entirety herein). Inone embodiment an effective amount of the isolated Compound 2 Form B iscombined in a single dosage form with the Syk inhibitor.

In one embodiment, the at least one additional chemotherapeutic agent isa protein cell death-1 (PD-1) inhibitor. PD-1 inhibitors are known inthe art, and include, for example, nivolumab (BMS), pembrolizumab(Merck), pidilizumab (CureTech/Teva), AMP-244 (Amplimmune/GSK),BMS-936559 (BMS), and MEDI4736 (Roche/Genentech). In one embodiment, aneffective amount of the isolated Compound 2 Form B is combined in asingle dosage form with the PD-1 inhibitor.

In an alternative embodiment isolated Compound 2 morphic Form A or D canbe used in an effective amount alone or in combination with anothercompound of the present invention or another bioactive agent to treat ahost such as a human with a disorder as described herein.

In one embodiment, the at least one additional chemotherapeutic agent isa B-cell lymphoma 2 (Bcl-2) protein inhibitor. BCL-2 inhibitors areknown in the art, and include, for example, ABT-199(4-[4-[[2-(4-Chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide),ABT-737(4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide), ABT-263((R)-4-(4-((4′-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide),GX15-070 (obatoclax mesylate,(2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4-methoxypyrrol-2-ylidene]indole;methanesulfonic acid))), 2-methoxy-antimycin A3, YC137(4-(4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester),pogosin, ethyl2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate,Nilotinib-d3, TW-37(N-[4-[[2-(1,1-Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl]benzamide),Apogossypolone (ApoG2), or G3139 (Oblimersen). In one embodiment, aneffective amount of the isolated Compound 2 Form B is combined in asingle dosage form with the at least one BCL-2 inhibitor.

In one embodiment, a combination described herein can be furthercombined with an additional therapeutic to treat the cancer. The secondtherapy can be an immunotherapy. As discussed in more detail below, aneffective amount of the isolated Compound 2 Form B can be conjugated toan antibody, radioactive agent, or other targeting agent that directsthe compound to the diseased or abnormally proliferating cell. Inanother embodiment, the combination is used in combination with anotherpharmaceutical or a biologic agent (for example an antibody) to increasethe efficacy of treatment with a combined or a synergistic approach. Inan embodiment, combination can be used with T-cell vaccination, whichtypically involves immunization with inactivated autoreactive T cells toeliminate a cancer cell population as described herein. In anotherembodiment, the combination is used in combination with a bispecificT-cell Engager (BiTE), which is an antibody designed to simultaneouslybind to specific antigens on endogenous T cells and cancer cells asdescribed herein, linking the two types of cells.

In one embodiment, the bioactive agent is a MEK inhibitor. MEKinhibitors are well known, and include, for example,trametinib/GSK1120212(N-(3-{3-Cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H-yl}phenyl)acetamide),selumetinib(6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide),pimasertib/AS703026NISC 1935369((S)—N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide),XL-518/GDC-0973(1-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidin-2-yl]azetidin-3-ol),refametinib/BAY869766/RDEA1 19(N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide),PD-0325901(N-[(2R)-2,3-Dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide),TAK733((R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione),MEK162/ARRY438162(5-[(4-Bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6-carboxamide),R05126766 (3-[[3-Fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidin-2-yloxychromen-2-one),WX-554, R04987655/CH4987655(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxazinan-2yl)methyl)benzamide),or AZD8330 (2-((2-fluoro-4-iodophenyl)amino)-N-(2hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide),U0126-EtOH, PD184352 (CI-1040), GDC-0623, BI-847325, cobimetinib,PD98059, BIX 02189, BIX 02188, binimetinib, SL-327, TAK-733, PD318088.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of binimetinib forthe treatment of melanoma, including BRAF-mutant melanoma andNRAS-mutant melanoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of cobimetinib(Cotellic) for the treatment of melanoma, including BRAF-mutant melanomaand NRAS-mutant melanoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of binimetinib forthe treatment of ovarian cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of selumetinib forthe treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of selumetinib forthe treatment of thyroid cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of trametinib(Mekinist) for the treatment of thyroid cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of trametinib(Mekinist) for the treatment of melanoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of trametinib(Mekinist) for the treatment of non-small cell lung cancer.

In one embodiment, the bioactive agent is a Raf inhibitor. Rafinhibitors are known and include, for example, Vemurafinib(N-[3-[[5-(4-Chlorophenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl]-1-propanesulfonamide),sorafenib tosylate(4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methylpyridine-2-carboxamide;4-methylbenzenesulfonate), AZ628(3-(2-cyanopropan-2-yl)-N-(4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)phenyl)benzamide),NVP-BHG712(4-methyl-3-(1-methyl-6-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino)-N-(3-(trifluoromethyl)phenyl)benzamide),RAF-265(1-methyl-5-[2-[5-(trifluoromethyl)-1H-imidazol-2-yl]pyridin-4-yl]oxy-N-[4-(trifluoromethyl)phenyl]benzimidazol-2-amine),2-Bromoaldisine(2-Bromo-6,7-dihydro-1H,5H-pyrrolo[2,3-c]azepine-4,8-dione), Raf KinaseInhibitor IV(2-chloro-5-(2-phenyl-5-(pyridin-4-yl)-1H-imidazol-4-yl)phenol),Sorafenib N-Oxide (4-[4-[[[[4-Chloro-3(trifluoroMethyl)phenyl]aMino]carbonyl]aMino]phenoxy]-N-Methyl-2pyridinecarboxaMide1-Oxide), PLX-4720, dabrafenib (GSK2118436), GDC-0879, RAF265, AZ 628,SB590885, ZM336372, GW5074, TAK-632, CEP-32496, LY3009120, and GX818(Encorafenib).

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of dabrafenib(Tafinlar) for the treatment of thyroid cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of dabrafenib(Tafinlar) for the treatment of melanoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of dabrafenib(Tafinlar) for the treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of encorafenib forthe treatment of melanoma, including BRAF-mutant melanoma.

In one embodiment, the additional therapy is a monoclonal antibody(MAb). Some MAbs stimulate an immune response that destroys cancercells. Similar to the antibodies produced naturally by B cells, theseMAbs “coat” the cancer cell surface, triggering its destruction by theimmune system. For example, bevacizumab targets vascular endothelialgrowth factor (VEGF), a protein secreted by tumor cells and other cellsin the tumor's microenvironment that promotes the development of tumorblood vessels. When bound to bevacizumab, VEGF cannot interact with itscellular receptor, preventing the signaling that leads to the growth ofnew blood vessels. Similarly, cetuximab and panitumumab target theepidermal growth factor receptor (EGFR), and trastuzumab targets thehuman epidermal growth factor receptor 2 (HER-2). MAbs that bind to cellsurface growth factor receptors prevent the targeted receptors fromsending their normal growth-promoting signals. They may also triggerapoptosis and activate the immune system to destroy tumor cells.

Another group of cancer therapeutic MAbs are the immunoconjugates. TheseMAbs, which are sometimes called immunotoxins or antibody-drugconjugates, consist of an antibody attached to a cell-killing substance,such as a plant or bacterial toxin, a chemotherapy drug, or aradioactive molecule. The antibody latches onto its specific antigen onthe surface of a cancer cell, and the cell-killing substance is taken upby the cell. FDA-approved conjugated MAbs that work this way includeado-trastuzumab emtansine, which targets the HER-2 molecule to deliverthe drug DM1, which inhibits cell proliferation, to HER-2 expressingmetastatic breast cancer cells.

Immunotherapies with T cells engineered to recognize cancer cells viabispecific antibodies (bsAbs) or chimeric antigen receptors (CARs) areapproaches with potential to ablate both dividing and non/slow-dividingsubpopulations of cancer cells.

Bispecific antibodies, by simultaneously recognizing target antigen andan activating receptor on the surface of an immune effector cell, offeran opportunity to redirect immune effector cells to kill cancer cells.The other approach is the generation of chimeric antigen receptors byfusing extracellular antibodies to intracellular signaling domains.Chimeric antigen receptor-engineered T cells are able to specificallykill tumor cells in a MHC-independent way.

In some embodiments, the combination can be administered to the subjectin further combination with other chemotherapeutic agents. Ifconvenient, the combination described herein can be administered at thesame time as another chemotherapeutic agent, in order to simplify thetreatment regimen. In some embodiments, the combination and the otherchemotherapeutic can be provided in a single formulation. In oneembodiment, the use of the compounds described herein is combined in atherapeutic regime with other agents. Such agents may include, but arenot limited to, tamoxifen, midazolam, letrozole, bortezomib,anastrozole, goserelin, an mTOR inhibitor, a PI3 kinase inhibitors, dualmTOR-PI3K inhibitors, MEK inhibitors, RAS inhibitors, ALK inhibitors,HSP inhibitors (for example, HSP70 and HSP 90 inhibitors, or acombination thereof), BCL-2 inhibitors, apopototic inducing compounds,AKT inhibitors, including but not limited to, MK-2206, GSK690693,Perifosine, (KRX-0401), GDC-0068, Triciribine, AZD5363, Honokiol,PF-04691502, and ipatasertib, Miltefosine; PD-1 inhibitors including butnot limited to, Nivolumab, CT-011, MK-3475, BMS936558, and AMP-514 orFLT-3 inhibitors, including but not limited to, P406, Dovitinib,Quizartinib (AC220), Amuvatinib (MP-470), Tandutinib (MLN518),ENMD-2076, and KW-2449, or combinations thereof.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ipatasertib forthe treatment of breast cancer, including triple negative breast cancer.

In one embodiment, the bioactive agent is an mTOR inhibitor. Examples ofmTOR inhibitors include but are not limited to vistusertib and rapamycinand its analogs, everolimus (Afinitor), temsirolimus, ridaforolimus,sirolimus, and deforolimus. Examples of MEK inhibitors include but arenot limited to tametinib/GSK1120212(N-(3-{3-Cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H-yl}phenyl)acetamide),selumetinob(6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide),pimasertib/AS703026NISC1935369((S)—N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide),XL-518/GDC-0973(1-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidin-2-yl]azetidin-3-ol),refametinib/BAY869766/RDEA119(N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide),PD-0325901(N-[(2R)-2,3-Dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide),TAK733((R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3d]pyrimidine-4,7(3H, 8H)-dione), MEK162/ARRY438162(5-[(4-Bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6carboxamide), R05126766 (3-[[3-Fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidin-2-yloxychromen-2-one),WX-554, R04987655/CH4987655(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxazinan-2yl)methyl)benzamide), or AZD8330(2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide).

In one embodiment, the bioactive agent is a RAS inhibitor. Examples ofRAS inhibitors include but are not limited to Reolysin and siG12D LODER.

In one embodiment, the bioactive agent is an ALK inhibitor. Examples ofALK inhibitors include but are not limited to Crizotinib, AP26113, andLDK378.

In one embodiment, the bioactive agent is a HSP inhibitor. HSPinhibitors include but are not limited to Geldanamycin or17-N-Allylamino-17-demethoxygeldanamycin (17AAG), and Radicicol. In aparticular embodiment, a compound described herein is administered incombination with letrozole and/or tamoxifen. Other chemotherapeuticagents that can be used in combination with the compounds describedherein include, but are not limited to, chemotherapeutic agents that donot require cell cycle activity for their anti-neoplastic effect.

Additional bioactive compounds include, for example, everolimus,trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693,RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258,GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054,PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, anaurora kinase inhibitor, a PIK-1 modulator, an HDAC inhbitor, a c-METinhibitor, a PARP inhibitor, a Cdk inhibitor, an IGFR-TK inhibitor, ananti-HGF antibody, a focal adhesion kinase inhibitor, a Map kinase (mek)inhibitor, a VEGF trap antibody, pemetrexed, panitumumab, amrubicin,oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab,zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifene,oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601,ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO1001, IPdR₁ KRX-0402, lucanthone, LY317615, neuradiab, vitespan, Rta744, Sdx 102, talampanel, atrasentan, Xr 311, romidepsin, ADS-100380,sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine,doxorubicin, liposomal doxorubicin, 5′-deoxy-5-fluorouridine,vincristine, temozolomide, ZK-304709, seliciclib; PD0325901, AZD-6244,capecitabine, L-Glutamic acid,N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-,disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan,tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethylstilbestrol), estradiol, estrogen, conjugated estrogen,bevacizumab, IMC-1C11, CHIR-258);3-[5-(methylsulfonylpiperadinemethyl)-indolyl-quinolone, vatalanib,AG-013736, AVE-0005, goserelin acetate, leuprolide acetate, triptorelinpamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate,megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide,megestrol acetate, CP-724714; TAK-165, HKI-272, erlotinib, lapatanib,canertinib, ABX-EGF antibody, erbitux, EKB-569, PKI-166, GW-572016,Ionafarnib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoylanalide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248,sorafenib, KRN951, aminoglutethimide, amsacrine, anagrelide,L-asparaginase, Bacillus Calmette-Guerin (BCG) vaccine, adriamycin,bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil,cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine,dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine,fludrocortisone, fluoxymesterone, flutamide, gleevec, gemcitabine,hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole,lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna,methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide,oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer,procarbazine, raltitrexed, rituximab, streptozocin, teniposide,testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine,13-cis-retinoic acid, phenylalanine mustard, uracil mustard,estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosinearabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin,mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat,COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668,EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifene,idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab,denileukin diftitox, gefitinib, bortezimib, paclitaxel, cremophor-freepaclitaxel, docetaxel, epithilone B, BMS-247550, BMS-310705,droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA-923, arzoxifene,fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339,ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin,40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001,ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646,wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin,erythropoietin, granulocyte colony-stimulating factor, zolendronate,prednisone, cetuximab, granulocyte macrophage colony-stimulating factor,histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylatedinterferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase,lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane,alemtuzumab, all-transretinoic acid, ketoconazole, interleukin-2,megestrol, immune globulin, nitrogen mustard, methylprednisolone,ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine,bexarotene, tositumomab, arsenic trioxide, cortisone, editronate,mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase,strontium 89, casopitant, netupitant, an NK-1 receptor antagonist,palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide,lorazepam, alprazolam, haloperidol, droperidol, dronabinol,dexamethasone, methylprednisolone, prochlorperazine, grani setron,ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, aplatelet-derived growth factor receptor alpha (PDGFR-α) antibody,epoetin alfa, darbepoetin alfa and mixtures thereof.

In one embodiment, an effective amount of the isolated Compound 2 Form Bdescribed herein can be combined with a PARP inhibitor selected fromniraparib tosylate monohydrate (Zejula), olaparib (Lynparza), rucaparibcamsylate (Rubraca), and talazoparib.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of niraparibtosylate monohydrate (Zejula) for the treatment of abnormal tissue ofthe female reproductive system, including ovarian epithelial cancer orfallopian tube cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of niraparibtosylate monohydrate (Zejula) for the treatment of peritoneal cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of olaparib(Lynparza) for the treatment of abnormal tissue of the femalereproductive system, including breast cancer, ovarian cancer, ovarianepithelial cancer or fallopian tube cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of olaparib(Lynparza) for the treatment of BRAC1 or BRAC2-mutated breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of olaparib(Lynparza) for the treatment of HER2− breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of olaparib(Lynparza) for the treatment of peritoneal cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of rucaparibcamsylate (Rubraca) for the treatment of abnormal tissue of the femalereproductive system, including breast cancer, ovarian cancer, ovarianepithelial cancer or fallopian tube cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of rucaparibcamsylate (Rubraca) for the treatment of peritoneal cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of talazoparib forthe treatment of abnormal tissue of the female reproductive system,including breast cancer, ovarian cancer, ovarian epithelial cancer orfallopian tube cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of talazoparib forthe treatment of BRAC1 or BRAC2-mutated breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of olaratumab forthe treatment of soft tissue sarcoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of savolitinib forthe treatment of adenocarcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of savolitinib forthe treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of savolitinib forthe treatment of renal cell carcinoma. In one embodiment, an effectiveamount of Compound 2 Form B is administered in combination with aneffective amount of vistusertib for the treatment of advanced breastcancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of vistusertib forthe treatment of advanced breast cancer.

In one embodiment, an effective amount of the isolated Compound 2 Form Bdescribed herein can be combined with a chemotherapeutic selected from,but are not limited to, Imatinib mesylate (Gleevac®), Dasatinib(Sprycel®), Nilotinib (Tasigna®), Bosutinib (Bosulif®), Trastuzumab(Herceptin®), Pertuzumab (Perjeta™), Lapatinib (Tykerb®), Gefitinib(Iressa®), Erlotinib (Tarceva®), Cetuximab (Erbitux®), Panitumumab(Vectibix®), Vandetanib (Caprelsa®), Vemurafenib (Zelboraf®), Vorinostat(Zolinza®), Romidepsin (Istodax®), Bexarotene (Tagretin®), Alitretinoin(Panretin®), Tretinoin (Vesanoid®), Carfilizomib (Kyprolis™),Pralatrexate (Folotyn®), Bevacizumab (Avastin®), Ziv-aflibercept(Zaltrap®), Sorafenib (Nexavar®), Sunitinib (Sutent®), Pazopanib(Votrient®), Regorafenib (Stivarga®), and Cabozantinib (Cometriq™).

In one embodiment, an effective amount of the isolated Compound 2 Form Bdescribed herein can be combined with a CD4/6 inhibitor includingabemaciclib (Versenio), palbociclib (Ibrance), or trilaciclib.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of abemaciclib(Versenio) for the treatment of breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of abemaciclib(Versenio) for the treatment of HR+ HER2− breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of palbociclib(Ibrance) for the treatment of breast cancer. In one embodiment, aneffective amount of Compound 2 Form B is administered in combinationwith an effective amount of palbociclib (Ibrance) for the treatment ofHR+ HER2− breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of palbociclib(Ibrance) for the treatment of breast cancer. In one embodiment, aneffective amount of Compound 2 Form B is administered in combinationwith an effective amount of palbociclib (Ibrance) for the treatment ofmetastatic triple negative breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of palbociclib(Ibrance) for the treatment of small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of cabozantinibS-malate (Cometrig™) for the treatment of thyroid cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of cabozantinibS-maleate (Cometrig™) for the treatment of renal cell carcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of dasatinib(Sprycel) for the treatment of leukemia, including acute lymphoblasticleukemia or chronic myelogenous leukemia.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of dasatinib(Sprycel) for the treatment of prostate cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of Erlotinib(Tarceva®) for the treatment of prostate cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of Gefitinib(Iressa®) for the treatment of prostate cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of imatinibmesylate (Gleevec) for the treatment of leukemia, including acutelymphoblastic leukemia, chronic eosinophilic leukemia, hypereosinophilicsyndrome, or chronic myelogenous leukemia.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of trastuzumab(Herceptin) for the treatment of adenocarcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of trastuzumab(Herceptin) for the treatment of breast cancer, including HER2+ breastcancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of imatinibmesylate (Gleevec) for the treatment of tumors, including but notlimited to dermatofibrosarcoma protuberans and gastrointestinal stromaltumors.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of imatinibmesylate (Gleevec) for the treatment ofmyelodysplastic/myeloproliferative neoplasms.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of imatinibmesylate (Gleevec) for the treatment of systemic mastocytosis.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of nilotinib(Tasigna) for the treatment of chronic myelogenous leukemia, includingPhiladelphia chromosome positive chronic myeloid leukemia (Ph+ CML).

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of pazopanibhydrochloride (Votrient) for the treatment of renal cell carcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of pazopanibhydrochloride (Votrient) for the treatment of soft tissue sarcoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of regorafenib(Stivarga) for the treatment of colorectal cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of regorafenib(Stivarga) for the treatment of gastrointestinal stromal tumor.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of regorafenib(Stivarga) for the treatment of hepatocellular carcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of sorafenibTosylate (Nexavar) for the treatment of carcinoma, includinghepatocellular carcinoma or renal cell carcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of sunitinib malate(Sutent) for the treatment of gastrointestinal stromal tumor.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of sunitinib malate(Sutent) for the treatment of pancreatic cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of sunitinib malate(Sutent) for the treatment of renal cell carcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of vemurafenib(Zelboraf) for the treatment of Erdheim-Chester disease.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of vemurafenib(Zelboraf) for the treatment of melanoma.

In certain aspects, the additional therapeutic agent is ananti-inflammatory agent, a chemotherapeutic agent, a radiotherapeutic,additional therapeutic agents, or immunosuppressive agents.

Suitable chemotherapeutic agents include, but are not limited to,radioactive molecules, toxins, also referred to as cytotoxins orcytotoxic agents, which includes any agent that is detrimental to theviability of cells, agents, and liposomes or other vesicles containingchemotherapeutic compounds. General anticancer pharmaceutical agentsinclude: Vincristine (Oncovin®) or liposomal vincristine (Marqibo®),Daunorubicin (daunomycin or Cerubidine®) or doxorubicin (Adriamycin®),Cytarabine (cytosine arabinoside, ara-C, or Cytosar®), L-asparaginase(Elspar®) or PEG-L-asparaginase (pegaspargase or Oncaspar®), Etoposide(VP-16), Teniposide (Vumon®), 6-mercaptopurine (6-MP or Purinethol®),Methotrexate, Cyclophosphamide (Cytoxan®), Prednisone, Dexamethasone(Decadron), imatinib (Gleevec®), dasatinib (Sprycel®), nilotinib(Tasigna®), bosutinib (Bosulif®), and ponatinib (Iclusig™) Examples ofadditional suitable chemotherapeutic agents include but are not limitedto 1-dehydrotestosterone, 5-fluorouracil decarbazine, 6-mercaptopurine,6-thioguanine, actinomycin D, adriamycin, aldesleukin, alkylatingagents, allopurinol sodium, altretamine, amifostine, anastrozole,anthramycin (AMC)), anti-mitotic agents, cis-dichlorodiamine platinum(II) (DDP) cisplatin), diamino dichloro platinum, anthracycline, anantibiotic, an antimetabolite, asparaginase, BCG live (intravesical),betamethasone sodium phosphate and betamethasone acetate, bicalutamide,bleomycin sulfate, busulfan, calcium leucouorin, calicheamicin,capecitabine, carboplatin, lomustine (CCNU), carmustine (BSNU),Chlorambucil, Cisplatin, Cladribine, Colchicin, conjugated estrogens,Cyclophosphamide, Cyclothosphamide, Cytarabine, Cytarabine, cytochalasinB, Cytoxan, Dacarbazine, Dactinomycin, dactinomycin (formerlyactinomycin), daunirubicin HCl, daunorucbicin citrate, denileukindiftitox, Dexrazoxane, Dibromomannitol, dihydroxy anthracin dione,Docetaxel, dolasetron mesylate, doxorubicin HCl, dronabinol, E. coliL-asparaginase, emetine, epoetin-α, Erwinia L-asparaginase, esterifiedestrogens, estradiol, estramustine phosphate sodium, ethidium bromide,ethinyl estradiol, etidronate, etoposide citrororum factor, etoposidephosphate, filgrastim, floxuridine, fluconazole, fludarabine phosphate,fluorouracil, flutamide, folinic acid, gemcitabine HCl, glucocorticoids,goserelin acetate, gramicidin D, granisetron HCl, hydroxyurea,idarubicin HCl, ifosfamide, interferon α-2b, irinotecan HCl, letrozole,leucovorin calcium, leuprolide acetate, levamisole HCl, lidocaine,lomustine, maytansinoid, mechlorethamine HCl, medroxyprogesteroneacetate, megestrol acetate, melphalan HCl, mercaptipurine, mesna,methotrexate, methyltestosterone, mithramycin, mitomycin C, mitotane,mitoxantrone, nilutamide, octreotide acetate, ondansetron HCL,paclitaxel, pamidronate disodium, pentostatin, pilocarpine HCl,plimycin, polifeprosan 20 with carmustine implant, porfimer sodium,procaine, procarbazine HCl, propranolol, rituximab, sargramostim,streptozotocin, tamoxifen, taxol, teniposide, tenoposide, testolactone,tetracaine, thioepa chlorambucil, thioguanine, thiotepa, topotecan HCl,toremifene citrate, trastuzumab, tretinoin, valrubicin, vinblastinesulfate, vincristine sulfate, and vinorelbine tartrate.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of bosutinib(Bosulif®) for the treatment of chronic myelogenous leukemia (CML).

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ponatinibhydrochloride (Iclusig) for the treatment of leukemia, including acutelymphoblastic leukemia and chronic myelogenous leukemia.

Additional therapeutic agents that can be administered in combinationwith a compound disclosed herein can include bevacizumab, sutinib,sorafenib, 2-methoxyestradiol or 2ME2, finasunate, vatalanib,vandetanib, aflibercept, volociximab, etaracizumab (MEDI-522),cilengitide, erlotinib, cetuximab, panitumumab, gefitinib, trastuzumab,dovitinib, figitumumab, atacicept, rituximab, alemtuzumab, aldesleukine,atlizumab, tocilizumab, temsirolimus, everolimus, lucatumumab,dacetuzumab, HLL1, huN901-DM1, atiprimod, natalizumab, bortezomib,carfilzomib, marizomib, tanespimycin, saquinavir mesylate, ritonavir,nelfinavir mesylate, indinavir sulfate, belinostat, panobinostat,mapatumumab, lexatumumab, dulanermin, ABT-737, oblimersen, plitidepsin,talmapimod, P276-00, enzastaurin, tipifarnib, perifosine, imatinib,dasatinib, lenalidomide, thalidomide, simvastatin, celecoxib,bazedoxifene, AZD4547, rilotumumab, oxaliplatin (Eloxatin), PD0332991,ribociclib (LEE011), amebaciclib (LY2835219), HDM201, fulvestrant(Faslodex), exemestane (Aromasin), PIM447, ruxolitinib (INC424), BGJ398,necitumumab, pemetrexed (Alimta), and ramucirumab (IMC-1121B).

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of HR+, HER2-breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of pancreatic cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of gastrointestinal cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of renal cell carcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of everolimus(Afinitor) for the treatment of astrocytoma, including subependymalgiant cell astrocytoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of fulvestrant(Faslodex) for the treatment of breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of fulvestrant(Faslodex) for the treatment of HR+, HER2-breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ramucirumab forthe treatment of adenocarcinoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ramucirumab forthe treatment of non-small cell lung cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ramucirumab forthe treatment of colorectal cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ribociclib(Kisqali) for the treatment of breast cancer.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ribociclib(Kisqali) for the treatment of HR+ and HER2− breast cancer.

In one aspect of the present invention, a compound described herein canbe combined with at least one IDH1 or IDH2 inhibitor. In one embodiment,an effective amount of Compound 2 Form B is administered in combinationwith an effective amount of enasidenib mesylate (Idhifa) for thetreatment of acute myeloid leukemia.

In one aspect of the present invention, a compound described herein canbe combined with at least one fibroblast growth factor receptor (FGFR)tyrosine kinase inhibitor. In one embodiment, an effective amount ofCompound 2 Form B is administered in combination with an effectiveamount of erdafitinib for the treatment of urothelial cancer, includingmetastatic urothelial cancer.

In one aspect of the present invention, a compound described herein canbe combined with at least one ERK inhibitor.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of SCH772984 forthe treatment of melanoma, including BRAF-mutant melanoma or NRAS-mutantmelanoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ulixertinib forthe treatment of melanoma, including uveal melanoma.

In one embodiment, an effective amount of Compound 2 Form B isadministered in combination with an effective amount of ulixertinib forthe treatment of pancreatic cancer.

In one aspect of the present invention, a compound described herein canbe combined with at least one immunosuppressive agent. Theimmunosuppressive agent is preferably selected from the group consistingof a calcineurin inhibitor, e.g. a cyclosporin or an ascomycin, e.g.Cyclosporin A (NEORAL®), FK506 (tacrolimus), pimecrolimus, a mTORinhibitor, e.g. rapamycin or a derivative thereof, e.g. Sirolimus(RAPAMUNE®), Everolimus (Certican®), temsirolimus, zotarolimus,biolimus-7, biolimus-9, a rapalog, e.g., ridaforolimus, azathioprine,campath 1H, a S1P receptor modulator, e.g. fingolimod or an analoguethereof, an anti IL-8 antibody, mycophenolic acid or a salt thereof,e.g. sodium salt, or a prodrug thereof, e.g. Mycophenolate Mofetil(CELLCEPT®), OKT3 (ORTHOCLONE OKT3®), Prednisone, ATGAM®,THYMOGLOBULIN®, Brequinar Sodium, OKT4, T10B9.A-3A, 33B3.1,15-deoxyspergualin, tresperimus, Leflunomide ARAVA®, CTLAI-Ig,anti-CD25, anti-IL2R, Basiliximab (SIMULECT®), Daclizumab (ZENAPAX®),mizorbine, methotrexate, dexamethasone, ISAtx-247, SDZ ASM 981(pimecrolimus, Elidel®), CTLA41g (Abatacept), belatacept, LFA31g,etanercept (sold as Enbrel® by Immunex), adalimumab (Humira®),infliximab (Remicade®), an anti-LFA-1 antibody, natalizumab (Antegren®),Enlimomab, gavilimomab, antithymocyte immunoglobulin, siplizumab,Alefacept efalizumab, pentasa, mesalazine, asacol, codeine phosphate,benorylate, fenbufen, naprosyn, diclofenac, etodolac and indomethacin,aspirin and ibuprofen.

In certain embodiments, a compound described herein is administered tothe subject prior to treatment with another chemotherapeutic agent,during treatment with another chemotherapeutic agent, afteradministration of another chemotherapeutic agent, or a combinationthereof.

In some embodiments, an effective amount of the isolated Compound 2 FormB can be administered to the subject such that the otherchemotherapeutic agent can be administered either at higher doses(increased chemotherapeutic dose intensity) or more frequently(increased chemotherapeutic dose density). Dose-dense chemotherapy is achemotherapy treatment plan in which drugs are given with less timebetween treatments than in a standard chemotherapy treatment plan.Chemotherapy dose intensity represents unit dose of chemotherapyadministered per unit time. Dose intensity can be increased or decreasedthrough altering dose administered, time interval of administration, orboth.

In one embodiment of the invention, the compounds described herein canbe administered in a concerted regimen with another agent such as anon-DNA-damaging, targeted anti-neoplastic agent or a hematopoieticgrowth factor agent. It has been recently been reported that theuntimely administration of hematopoietic growth factors can have seriousside effects. For example, the use of the EPO family of growth factorshas been associated with arterial hypertension, cerebral convulsions,hypertensive encephalopathy, thromboembolism, iron deficiency, influenzalike syndromes and venous thrombosis. The G-CSF family of growth factorshas been associated with spleen enlargement and rupture, respiratorydistress syndrome, allergic reactions and sickle cell complications. Assuch, in one embodiment, the use of the compounds or methods describedherein is combined with the use of hematopoietic growth factorsincluding, but not limited to, granulocyte colony stimulating factor(G-CSF, for example, sold as Neupogen (filgrastin), Neulasta(peg-filgrastin), or lenograstin), granulocyte-macrophage colonystimulating factor (GM-CSF, for example sold as molgramostim andsargramostim (Leukine)), M-CSF (macrophage colony stimulating factor),thrombopoietin (megakaryocyte growth development factor (MGDF), forexample sold as Romiplostim and Eltrombopag) interleukin (IL)-12,interleukin-3, interleukin-11 (adipogenesis inhibiting factor oroprelvekin), SCF (stem cell factor, steel factor, kit-ligand, or KL) anderythropoietin (EPO), and their derivatives (sold as for exampleepoetin-α as Darbopoetin, Epocept, Nanokine, Epofit, Epogin, Eprex andProcrit; epoetin-β sold as for example NeoRecormon, Recormon andMicera), epoetin-delta (sold as for example Dynepo), epoetin-omega (soldas for example Epomax), epoetin zeta (sold as for example Silapo andReacrit) as well as for example Epocept, EPOTrust, Erypro Safe,Repoeitin, Vintor, Epofit, Erykine, Wepox, Espogen, Relipoeitin,Shanpoietin, Zyrop and EPIAO). In one embodiment, an effective amount ofthe isolated Compound 2 Form B is administered prior to administrationof the hematopoietic growth factor. In one embodiment, the hematopoieticgrowth factor administration is timed so that the compound's effect onHSPCs has dissipated. In one embodiment, the growth factor isadministered at least 20 hours after the administration of a compounddescribed herein.

If desired, multiple doses of a compound described herein can beadministered to the subject. Alternatively, the subject can be given asingle dose of a compound described herein. In one aspect of theinvention, a compound disclosed herein can be beneficially administeredin combination with any therapeutic regimen entailing radiotherapy,chemotherapy, or other therapeutic agents. In additional embodiments thecompounds disclosed herein can be beneficially administered incombination with therapeutic agents targeting auto-immune disorders.

In an alternative embodiment Compound 2 Form A, C, D, E, G, or H, isadministered in a combination described above instead of Compound 2 FormB to treat a host, typically a human, with a selected cancer, tumor,hyperproliferative condition or an inflammatory or immune disorder.

EXAMPLES Example 1. Conversion of Compound 1 to its HCl Counterpart,Compound 2

A representative synthesis of Compound 2 is provided in Scheme 1.

Compound 1 (0.9 kg. 1.9 moles, 1 eq) was charged to a 22 L flask anddissolved in aqueous, 2 M hydrochloric acid solution (3.78 L). Thesolution was heated to 50±5° C., stirred for 30 minutes, and theresulting mixture filtered over Celite (alternatively the solution maybe filtered through a 0.45 micron in-line filter) to afford Compound 2.The flask was rinsed with 0.1 M hydrochloric acid solution to collectany additional Compound 2. Compound 2 was then heated to 50±5° C. whileacetone (6.44 L) was slowly added. The solution was stirred at 50±5° C.for 30 minutes, the temperature was decreased to 20±5° C., and stirringcontinued for 2 hours. The solids were collected by filtration, washedwith acetone, and dried to afford 820.90 g of Compound 2 (82.1% yield).In one embodiment instead of acetone, ethanol is used.

Example 2. Morphic Forms of Compound 2

Eleven unique XRPD patterns (Form A-Form K) of Compound 2 were obtainedfrom crystallization and slurry experiments using various solvents. Theconditions and XRPD results for these crystallization experiments aregiven in Tables 1-4. Single solvent crystallizations (Table 1) resultedin weak crystalline forms or Form A. Binary solvent crystallizationsusing water (Table 2) and MeOH (Table 3) as the primary solvent resultedin weak crystalline forms and Form A, Form B, Form F, Form G, and FormH. Solids recovered from slurry experiments after one and seven days ofequilibration (Table 4) were analyzed by XRPD to determine thecrystalline form, and after seven days, Form A, Form B, Form C, Form D,and Form E were observed. FIG. 1 shows the XRPD patterns of Form A, FormB, and Form C. FIG. 2 shows the XRPD patterns of Form D, Form E, andForm F. FIG. 3 shows the XRPD patterns of Form G and Form H.

TABLE 1 Single Solvent Crystallization Conditions and Results VolumeTemp. Precipitation/ Solvent (mL) (° C.) Cooling Isolation XRPD Water2.0 60 Slow (20° C./hr) Turbid/Evap. Weak crystalline MeOH 0.5 60 Slow(20° C./hr) ppt/filter A EtOH 4.0 60 Slow (20° C./hr) ppt/filter A1-PrOH 4.0 60 Slow (20° C./hr) ppt/filter Weak crystalline 1-BuOH 4.0 60Slow (20° C./hr) ppt/filter A Water 2.0 60 Fast Cooling Turbid/Evap.Weak (4° C.) crystalline MeOH 0.5 60 Fast Cooling ppt/filter Weak (4°C.) crystalline EtOH 4.0 60 Fast Cooling ppt/filter A (4° C.) 1-PrOH 4.060 Fast Cooling ppt/filter Weak (4° C.) crystalline 1-BuOH 4.0 60 FastCooling ppt/filter Weak (4° C.) crystalline

TABLE 2 Binary Solvent Crystallizations using water as Primary SolventPrimary Anti Solvent/Vol. Temp. Solvent/Vol. Precipitation/ (mL) (° C.)Cooling (mL) Isolation XRPD Water/0.5 60.0 Fast Cooling (4° C.) EtOH/5.0Clear/Evap. Weak crystalline Water/0.5 60.0 Fast Cooling (4° C.)n-PrOH/5.0 Clear/Evap. Weak crystalline Water/0.5 60.0 Fast Cooling (4°C.) IPA/5.0 ppt/filter G Water/0.5 60.0 Fast Cooling (4° C.) MeCN/5.0ppt/filter Weak crystalline Water/0.5 60.0 Fast Cooling (4° C.) THF/3.0ppt/filter Weak crystalline Water/0.5 60.0 Fast Cooling (4° C.)Acetone/3.5 ppt/filter G Water/0.5 60.0 Slow Cooling EtOH/5.0Clear/Evap. Weak crystalline (20° C./hr) Water/0.5 60.0 Slow Coolingn-PrOH/5.0 Clear/Evap. H (20° C./hr) Water/0.5 60.0 Slow Cooling IPA/5.0ppt/filter B (20° C./hr) Water/0.5 60.0 Slow Cooling MeCN/5.0 ppt/filterA (20° C./hr) Water/0.5 60.0 Slow Cooling THF/3.0 ppt/filter G (20°C./hr) Water/0.5 60.0 Slow Cooling Acetone/3.5 ppt/filter B (20° C./hr)

TABLE 3 Binary Solvent Crystallizations using MeOH as Primary SolventPrimary Anti Solvent/Vol. Temp. Solvent/Vol. Precipitation/ (mL) (° C.)Cooling (mL) Isolation XRPD MeOH/0.5 60.0 Fast Cooling (4° C.) EtOH/5.0ppt/filter A MeOH/0.5 60.0 Fast Cooling (4° C.) n-PrOH/5.0 ppt/filterWeak crystalline MeOH/0.5 60.0 Fast Cooling (4° C.) IPA/2.5 ppt/filter FMeOH/0.5 60.0 Fast Cooling (4° C.) n-BuOH/5.0 ppt/filter Weakcrystalline MeOH/0.5 60.0 Fast Cooling (4° C.) MeCN/2.5 ppt/filter AMeOH/0.5 60.0 Fast Cooling (4° C.) THF/0.5 ppt/filter A MeOH/0.5 60.0Fast Cooling (4° C.) 2-MeTHF/0.1 ppt/filter A MeOH/0.5 60.0 Fast Cooling(4° C.) EtOAc/0.2 ppt/filter Weak crystalline MeOH/0.5 60.0 Fast Cooling(4° C.) IPAc/0.1 ppt/filter A MeOH/0.5 60.0 Fast Cooling (4° C.)Acetone/0.5 ppt/filter A MeOH/0.5 60.0 Slow Cooling MEK/0.2 ppt/filter A(20° C./hr) MeOH/0.5 60.0 Slow Cooling MIBK/0.1 ppt/filter Weakcrystalline (20° C./hr) MeOH/0.5 60.0 Slow Cooling DCM/5.0 Clear/Evap. A(20° C./hr) MeOH/0.5 60.0 Slow Cooling Toluene/1.5 ppt/filter A (20°C./hr) MeOH/0.5 60.0 Slow Cooling MTBE/0.1 ppt/filter A (20° C./hr)MeOH/0.5 60.0 Slow Cooling EtOH/5.0 ppt/filter Weak crystalline (20°C./hr) MeOH/0.5 60.0 Slow Cooling n-PrOH/5.0 ppt/filter Weak crystalline(20° C./hr) MeOH/0.5 60.0 Slow Cooling IPA/2.5 ppt/filter A (20° C./hr)MeOH/0.5 60.0 Slow Cooling n-BuOH/5.0 ppt/filter Weak crystalline (20°C./hr) MeOH/0.5 60.0 Slow Cooling MeCN/2.5 ppt/filter Weak crystalline(20° C./hr) MeOH/0.5 60.0 Slow Cooling THF/0.5 ppt/filter Weakcrystalline (20° C./hr) MeOH/0.5 60.0 Slow Cooling 2-MeTHF/0.1ppt/filter A (20° C./hr) MeOH/0.5 60.0 Slow Cooling EtOAc/0.2 ppt/filterA (20° C./hr) MeOH/0.5 60.0 Slow Cooling IPAc/0.1 ppt/filter A (20°C./hr) MeOH/0.5 60.0 Slow Cooling Acetone/0.5 ppt/filter Weakcrystalline (20° C./hr) MeOH/0.5 60.0 Slow Cooling MEK/0.2 ppt/filter A(20° C./hr) MeOH/0.5 60.0 Slow Cooling MIBK/0.1 ppt/filter A (20° C./hr)MeOH/0.5 60.0 Slow Cooling DCM/5.0 Clear/Evap. A (20° C./hr) MeOH/0.560.0 Slow Cooling Toluene/1.5 ppt/filter Weak crystalline (20° C./hr)MeOH/0.5 60.0 Slow Cooling MTBE/0.1 ppt/filter A (20° C./hr)

TABLE 4 Slurry Experiments of Compound 2 Solvent Time point Time pointVol. (1 day) (7 days) Solvent (mL) Method XRPD XRPD IPA 1.0 Stirring atRT A F MeCN 1.0 Stirring at RT D D THF 1.0 Stirring at RT WeakCrystalline E 2-MeTHF 1.0 Stirring at RT Weak Crystalline B EtOAc 1.0Stirring at RT A C IPAc 1.0 Stirring at RT A with extra peak B Acetone1.0 Stirring at RT E B MEK 1.0 Stirring at RT Weak Crystalline B MIBK1.0 Stirring at RT E B Toluene 1.0 Stirring at RT E B MTBE 1.0 Stirringat RT A B n-Heptane 1.0 Stirring at RT A A c-Hexane 1.0 Stirring at RT AA

Example 2. Characterization of Compound 2 Morphic Forms

A summary of characterization data of all isolated forms of Compound 2is given in Table 5. Forms A, B, and D were evaluated as solid stateforms.

TABLE 5 Characterization Data of Morphic Forms of Compound 2 XRPDPossible 1H NMR % Cl Pattern Form DSC (° C.) TGA (wt loss) (DMSO-d₆)(API:HCl) A Hydrate Endotherms at Onset 5.7 wt % loss at 66.0° C.,Contains 11.1% 110.3, Onset 5.4 wt % loss at water (1:1.67)* 275.6,344.8 215.5° C., Onset 6.2 wt % loss at 314.0° C. B Hydrate Endothermsat Onset 5.1 wt % loss at 60.9° C., Contains 11.90% 105.2, Onset 7.2 wt% loss at water and (1:1.81) 220.8, 265.6, 198.3° C., Onset 7.8 wt %loss residual 350.6 at 319.6° C. solvent C EtOAc Endotherms at Onset 1.6wt % loss at 72.9° C., Contains Not solvate 95.1, Onset 5.1 wt % loss atwater and determined 235.6, 257.8, 192.0° C., Onset 0.9 wt % loss EtOAcas 344.6 at 223.4° C., Onset 6.9 wt % residual loss at 306.7° C. solventD Hydrate Endotherms at Onset 6.0 wt % loss at 68.8° C., Contains 12.23%108.3, Onset 6.0 wt % loss at water and (1:1.87) 266.1, 347.0 207.6° C.,Onset 3.6 wt % loss residual at 304.9° C., Onset 6.6 wt % solvent lossat 324.7° C. E Acetone Endotherms at Onset 1.0 wt % loss at 41.9° C.,Contains Not solvate 70.3, Onset 1.1 wt % loss at 61.5° C., water anddetermined 275.2, 345.9 Onset 1.0 wt % loss at 93.2° C., acetone asExotherm at Onset 5.0 wt % loss at residual 220.0 211.6° C., Onset 5.6wt % loss solvent at 308.5° C. F Unstable Endotherms at Onset 8.0 wt %loss at 43.7° C., Contains Not hydrate 73.2, Onset 2.1 wt % loss atwater determined 214.5, 303.4, 190.7° C., Onset 7.6 wt % loss 329.7 at308.8° C. Exotherm at 277.8 G Anhydrate Endotherms at Onset 4.5 wt %loss at 47.2° C., Contains Not 81.8, Onset 3.1 wt % loss at 86.6° C.,water determined 120.8, 268.2, Onset 4.5 wt % loss at 347.9 213.3° C.,Onset 4.6 wt % loss at 311.2° C. H n-PrOH Endotherms at Onset 1.9 wt %loss at 45.6° C., Contains Not solvate 110.5, Onset 4.6 wt % loss at71.9° C., water and n- determined 225.6, 274.5, Onset 1.8 wt % loss atPrOH as 346.3 187.9° C., Onset 2.2 wt % loss residual at 222.1° C.,Onset 3.0 wt % solvent loss at 303.0° C., Onset 2.2 wt % loss at 325.2°C.

In one embodiment Form A is characterized by at least one XRPD peaks at7.4±0.2°, 9.0±0.2°, or 12.3±0.2° 2theta. In one embodiment Form B ischaracterized by at least one XRPD peaks at 6.4±0.2°, or 9.5±0.2°2theta. In one embodiment Form C is characterized by at least one XRPDpeaks at 5.3±0.2°, or 7.2±0.2° 2theta. In one embodiment Form D ischaracterized by at least one XRPD peaks at 5.6±0.2°, or 8.2±0.2°2theta. In one embodiment Form E is characterized by at least one XRPDpeak at 5.5±0.2°, or 6.7±0.2° 2theta. In one embodiment Form E ischaracterized by at least one XRPD peak at 5.5±0.2°, or 6.7±0.2° 2theta.In one embodiment Form F is characterized by a XRPD peak at 7.2±0.2°2theta. In one embodiment Form G is characterized by a XRPD peak at6.7±0.2° 2theta. In one embodiment Form H is characterized by a XRPDpeak at 6.6±0.2° 2theta.

Example 3. Dynamic Vapor Sorption Experiments of Form a, Form B, andForm D

Dynamic vapor sorption experiments were performed on Form A, Form B, andForm D. Table 6 provides the results of the DVS experiment.

TABLE 6 Moisture Sorption Data of Forms A, B, and D XRPD % wt change %wt change XRPD (pre DVS) at 60% RH at 90% RH (post DVS) Form A 14.9 15.8Form K Form B 5.8 5.9 Form B Form D 4.4 17.0 Form K

Form A was found to be unstable in the moisture sorption experiment. Thematerial adsorbed 14.9 wt % moisture at 60% RH and 15.8 wt % at 90% RH.After the moisture sorption experiment, the sample was dried at 60° C.and 0% RH and the result of the XRPD analysis of dried sample showed anew Form (Form K). The DVS analysis of Form A is shown in FIG. 4A. FormD was also found to be unstable in the moisture sorption experiment. Thematerial adsorbed 4.4 wt % moisture at 60% RH and 17.0 wt % at 90% RH.After the moisture sorption experiment, the sample was dried at 60° C.and 0% RH and the result of the XRPD analysis of dried sample showedForm K. The DVS analysis of Form D is shown in FIG. 4B.

Unlike Form A and Form D, Form B was stable in the moisture sorptionexperiment. The material adsorbed 5.8 wt % moisture at 60% RH and 5.9 wt% at 90% RH. After drying at 60° C. and 0% RH for two hours, the XRPDpattern remained unchanged as Form B. The DVS analysis of Form B isshown in FIG. 4C.

FIG. 5A is a comparison of the XRPD pattern of Form A before DVSanalysis and the new pattern (Form K) that resulted from DVS. FIG. 5B isa comparison of the XRPD pattern of Form D before DVS and the pattern(Form K) that resulted after DVS.

Example 4. Stability Study of Forms a, B, and D Under Thermal Stress

Forms A, B, and D were stored in an oven maintained at 60° C. for 7days. No change in the XRPD pattern was observed for Form B or Form D. Anew pattern was found for Form A at the conclusion of the stabilitystudy, however after equilibrium for three days at room temperature, theXRPD of the new Form revealed that it had converted back to Form A. FIG.6 compares the XRPD patterns of Form A, Form B, and Form D to referencematerial. FIG. 6 also shows the new pattern that resulted from exposingForm A to thermal stress along with the Form A pattern that resultedafter three additional days at room temperature.

Example 5. Recrystallization Procedures to Produce Form B from Compound2

Recrystallization studies were conducted to define a procedure toimprove chromatographic purity. All recrystallization procedures inTable 7 involved dissolving Compound 2 in concentrated HCl and thenadding the anti-solvent, acetone. The differences in the processes aresubtle but important in terms of their results.

Recrystallization Process 1: Compound 1 was charged to an appropriatelysized flask or reactor, dissolved in aqueous hydrochloric acid solutionand heated to at least 55±10° C. The solution was stirred for about 45minutes and the resulting mixture was filtered through an in-linefilter. Acetone was added at 55±10° C. over the course of an hour andthe solution was stirred for about an additional hour. The temperaturewas decreased to about 25±5° C., and the solution was stirred for atleast 2 hours. The solids were collected by filtration, washed withacetone, and dried to afford Compound 2 form B.

Recrystallization Process 2: Compound 1 was charged to an appropriatelysized flask or reactor, dissolved in aqueous hydrochloric acid solutionand heated to at least 55±10° C. The solution was stirred for about 45minutes and the resulting mixture was filtered through an in-linefilter. The temperature was decreased to about 25±5° C., and thesolution was stirred for at least 2 hours. Acetone was added at 25±5° C.over the course of an hour and the solution was stirred for anadditional two hours. The solids were collected by filtration, washedwith acetone, and dried to afford Compound 2 form D.

Recrystallization Process 3: Compound 1 was charged to an appropriatelysized flask or reactor, dissolved in aqueous hydrochloric acid solutionand heated to at least 55±10° C. The solution was stirred for about 45minutes and the resulting mixture was filtered through an in-linefilter. The temperature was decreased to about 25±5° C. and the solutionwas stirred for at least 2 hours. The solids were collected byfiltration, washed with acetone, and dried to afford Compound 2 form D.

TABLE 7 Effect of crystallization procedures on purging ofchromatographic impurities from Compound 1 Recrys Recrys Recrys Process1 Process 2 Process 3 RRT % area % area % area % area 1.11 1.13 1.110.87 0.27 1.37 0.14 0.15 0.13 ND 1.62 0.14 ND 0.13 ND

While conducting the experiments presented in Table 7, it was discoveredthat not all recrystallization processes resulted in the preferred solidstate form, Form B. Specifically, Recrystallization Processes 2 and 3result in a different solid state form (putative Form D) whereasRecrystallization 1 reproducibly provides Form B. In one embodiment,Compound 2 is converted to Form D by Recrystallization Procedure 2 and 3and Form D is converted to Form B by Recrystallization Process 1.

Example 6. XRPD Analysis of Compound 2, Morphic Form B

The XRPD pattern of Form B was collected with a PANalytical X'Pert PROMPD diffractometer using an incident beam of Cu radiation produced usingan Optix long, fine-focus source. An elliptically graded multilayermirror was used to focus Cu Kα X-rays through the specimens and onto thedetector. Prior to the analysis, a silicon specimen (NIST SRM 640e) wasanalyzed to verify the observed position of the Si 111 peak isconsistent with the NIST-certified position. The sample was sandwichedbetween 3-μm-thick films and analyzed in transmission geometry. Abeamstop, short anti-scatter extension and an anti-scatter knife edgewere used to minimize the background generated by air. Soller slits forthe incident and diffracted beams were used to minimize broadening fromaxial divergence. The diffraction patterns were collected using ascanning position-sensitive detector (X'Celerator) located 240 mm fromthe specimens and Data Collector software v. 2.2b. Data acquisitionparameters for each pattern are displayed above the image in the Datasection of this report including the divergence slit (DS) before themirror.

The XRPD pattern of pure Form B along with the indexing solution isshown in FIG. 7 . The pure Form B XRPD pattern exhibited sharp peaks,indicating the sample was composed of crystalline material. The allowedpeak positions from the XRPD indexing solution are 6.5, 8.1, 9.4, 9.6,10.2, 10.6, 11.2, 12.2, 12.9, 13.0, 13.3, 13.4, 14.0, 14.4, 14.6, 15.0,15.9, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.6, 18.9, 19.1,19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 20.6, 21.3, 21.4,21.8, 22.0, 22.2, 22.3, 22.4, 22.5, 22.8, 23.0, 23.1, 23.4, 23.8, 24.1,24.2, 24.3, 24.4, 24.5, 24.6, 25.4, 25.6, 25.7, 25.9, 26.0, 26.1, 26.3,26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.2, 27.3, 27.5, 27.6, 27.7, 27.9,28.3, 28.4, 28.5, 28.7, 28.9, 29.0, 29.1, 29.3, 29.4, 29.5, 29.6, 29.7,29.8, 29.9, 30.0, 30.3, 30.4, 30.5, 30.6, 30.7, 30.9, 31.2, 31.5, 31.6,31.7, 31.8, 31.9, 32.0, 32.2, 32.3, 32.4, 32.5, 32.6, 32.7, 32.8, 33.1,33.2, 33.3, 33.6, 33.7, 33.8, 34.0, 34.1, 34.2, 34.3, 34.6, 34.7, 34.8,35.0 35.2, 35.3, 35.5, 35.6, 35.9, 36.0, 36.2, 36.5, 36.6, 36.7, 36.8,36.9, 37.1, 37.2, 37.3, 37.4, 37.5, 37.6, 37.7, 37.8, 37.9, 38.2, 38.3,38.4, 38.5, 38.6, 38.7, 38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5,39.6, 39.7, 39.8, 39.9, and 40.0° 2θ.

For example, Form B's XRPD may be indexed as follows 6.47, 8.08, 9.42,9.59, 10.18, 10.62, 11.22, 12.17, 12.91, 12.97, 13.27, 13.37, 14.03,14.37, 14.63, 15.02, 15.93, 16.20, 16.35, 16.43, 16.47, 16.81, 18.10,18.35, 18.41, 18.50, 18.55, 18.6,0 18.91, 19.11, 19.15, 19.24, 19.34,19.43, 19.51, 19.61, 19.65, 19.76, 19.85, 19.90, 20.44, 20.61, 21.34,21.43, 21.84, 21.95, 22.17, 22.28, 22.30, 22.33, 22.44, 22.54, 22.76,22.81, 22.97, 23.00, 23.11, 23.42, 23.80, 24.11, 24.22, 24.34, 24.38,24.40, 24.48, 24.56, 24.57, 25.40, 25.56, 25.57, 25.59, 25.72, 25.74,25.94, 25.99, 26.11, 26.28, 26.29, 26.37, 26.51, 26.58, 26.61, 26.73,26.81, 26.92, 27.15, 27.19, 27.23, 27.31, 27.49, 27.57, 27.61, 27.71,27.88, 27.94, 28.27, 28.41, 28.53, 28.71, 28.74 28.86, 28.94, 28.98,29.03, 29.06, 29.08, 29.25, 29.30, 29.38, 29.51, 29.57, 29.61, 29.70,29.73, 29.75, 29.90, 29.95, 30.31, 30.38, 30.42, 30.54, 30.55, 30.66,30.73, 30.85, 30.87, 30.89, 31.23, 31.51, 31.55, 31.61, 31.70, 31.76,31.77, 31.80, 31.81, 31.82, 31.82, 31.90, 31.91, 31.95, 32.17, 32.21,32.23, 32.25, 32.36, 32.37, 32.43, 32.53, 32.54, 32.56, 32.61, 32.73,32.80, 32.82, 33.05, 33.13, 33.17, 33.22, 33.28, 33.30, 33.60, 33.65,33.71, 33.76, 33.77, 33.99, 34.01, 34.01, 34.05, 34.10, 34.17, 34.29,34.55, 34.60, 34.62, 34.63, 34.68, 34.75, 34.76, 35.03, 35.16, 35.19,35.21, 35.25, 35.31, 35.46, 35.61, 35.63, 35.85, 35.86, 35.90, 35.97,36.19, 36.45, 36.56, 36.58, 36.67, 36.68, 36.70, 36.71, 36.77, 36.85,36.87, 36.90, 37.09, 37.19, 37.27, 37.28, 37.29, 37.32, 37.33, 37.37,37.38, 37.48, 37.48, 37.50, 37.51, 37.54, 37.61, 37.64, 37.65, 37.68,37.69, 37.71, 37.74, 37.74, 37.76, 37.81, 37.83, 37.93, 37.94, 38.15,38.19, 38.32, 38.36, 38.39, 38.46, 38.59, 38.63, 38.69, 38.76, 38.79,38.85, 38.87, 38.88, 38.96, 38.98, 39.02, 39.05, 39.19, 39.27, 39.33,39.36, 39.39, 39.43, 39.44, 39.53, 39.53, 39.6, 39.61, 39.70, 39.71,39.72, 39.82, 39.87, 39.9, and 39.98° 2θ.

Observed peaks for Form B include 9.5±0.2, 18.1±0.2, 19.3±0.2, 22.4±0.2,26.6±0.2, and 27.7±0.2, ° 2θ.

Agreement between the allowed peak positions, marked with bars, and theobserved peaks indicated a consistent unit cell determination.Successful indexing of the pattern indicated that the sample wascomposed primarily of a single crystalline phase. Space groupsconsistent with the assigned extinction symbol, unit cell parameters,and derived quantities are given in Table 8.

TABLE 8 Parameters of the XRPD of Compound 2, Form B Bravais TypeC-centered Monoclinic a [Å] 27.719 b [Å] 9.796 c [Å] 22.221 α [deg] 90 β[deg] 100.16 γ [deg] 90 Volume [Å³/cell] 5,939.0 Chiral contents Notspecified Extinction Symbol C 1 c 1 Space Group(s) Cc (9), C2/c (15)

In one embodiment, Form B is characterized by an XRPD pattern comprisingat least two 2theta values selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°,14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, FormB is characterized by an XRPD pattern comprising at least three 2thetavalues selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°,18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, Form B ischaracterized by an XRPD pattern comprising at least four 2theta valuesselected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°,19.9±0.2°, and 22.4±0.2°. In one embodiment, Form B is characterized byan XRPD pattern comprising at least five 2theta values selected from6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and22.4±0.2°. In one embodiment, Form B is characterized by an XRPD patterncomprising at least six 2theta values selected from 6.5±0.2°, 9.5±0.2°,14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In oneembodiment, Form B is characterized by an XRPD pattern comprising the2theta values selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°,18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, Form B ischaracterized by an XRPD pattern comprising at least the 2theta value of9.5±0.4°.

Example 7. Six- and Twelve-Month Stability Study of Form B at 25° C./60%RH and at 40° C./75% RH Conditions

Form B was stored at 25° C./60% RH for twelve months and at 40° C./75%RH for six months.

Table 9 shows the results from the 25° C./60% RH storage conditions andTable 10 shows the results at the 40° C./75% RH. In both conditions,Form B was 99.5% pure and the XRPD spectrum conformed to the referencespectrum at the longest time point studied.

TABLE 9 Twelve-month Stability Study at the 25° C./60% RH ConditionsStorage Time Test 0 M 3 M 6 M 9 M 12 M Appearance Yellow Yellow YellowYellow Yellow solid solid solid solid solid Moisture  4.9% 4.07% 7.53%7.29% 7.48% (%) XRPD Form B N/A N/A N/A Form B HPLC 99.5% 99.5%  99.5% 99.6%  99.5%  Purity

TABLE 10 Six-month Stability Study at the 40° C./75% RH ConditionsStorage Time Test 0 M 1 M 3 M 6 M Appearance Yellow solid Yellow solidYellow solid Yellow solid Moisture (%)  4.9% 2.75% 7.47% 7.53% XRPD FormB N/A Form B Form B HPLC Purity 99.5% 99.5%  99.5%  99.5% 

Example 8. Conversion of Impure Form B Material to Pure Form B Material

Pure Form B was isolated from impure Form B, material that wascharacterized as containing a residual amount of an unknown form inaddition to Form B. The difference in the XRPD patterns of impure Form Band pure Form B is shown in FIG. 8 . (In the following experimentsdescribed below, pure Form B is the Form B as characterized in Example6.) The pattern of pure Form B is visually similar to the pattern ofimpure Form B, with the exception of the absence of peaks at 2-thetaangles of approximately 4.0 and 5.6°. As is common in XRPD analysis,there are also differences in relative peak intensities that are likelydue to preferred orientation and/or particle statistics effects.

Solubility studies, small scouting experiments, and experiments withdrying conditions were first performed to confirm the conditionssuitable for the conversion to pure Form B. TG-IR Characterization wasperformed on a number of isolated samples of Form B. Once conditionswere confirmed, the conversion from impure material to pure material wasconducted in water:acetone 1:2 (v/v) slurry at 125 mg/mL concentrationand 30° C. for 43 hours as described in more detail below.

Solubility Estimate Experiments for the Development of ConditionsSuitable for Recrystallization

Solubility estimates of impure Form B were attempted in variouspredominantly HCl acidic aqueous acetone solvent mixtures using analiquot addition method that involved visual observation. Aliquots ofvarious solvents or diluent/organic solvent mixtures were added tomeasured amounts of impure Form B with agitation (typically sonication)at ambient temperature until complete dissolution was achieved, asjudged by visual observation. Solubilities were calculated based on thetotal solvent used to give a solution; actual solubilities may begreater because of the volume of solvent portions utilized or a slowrate of dissolution. If dissolution did not occur as determined byvisual assessment, the value was reported as “<”. If dissolutionoccurred at the first aliquot, the value was reported as “>”. Due to thehaziness of the obtained samples, effective solubility estimates weredifficult to discern. In general, impure Form B showed very limitedsolubility (3-7 mg/mL) in the tested solvent mixtures (Table 11).

TABLE 11 Approximate Solubility of Impure Form B Solvent/ TemperatureSolubility^(a) Solvent System (° C.) (mg/mL) Observation 1M HCl:acetoneambient <1 solids remained 20:80 0.5M HCl:acetone ambient <1 solidsremained 20:80 0.1M HCl:acetone ambient <1 solids remained 20:80 1MHCl:acetone ambient <1 solids remained 10:90 0.5M HCl:acetone ambient <1solids remained 10:90 1M HCl:acetone ambient <1 solids remained 5:95 1MHCl:acetone ambient 2 hazy solution 33:67 1M HCl:acetone ambient 4 hazysolution 50:50 1M HCl:acetone ambient 4 hazy solution 60:40 1MHCl:acetone ambient 6 hazy solution 67:33 1.0M HCl:acetone ambient 7hazy solution 75:25 2.0M HCl ambient 4 hazy solution 5.0M HCl ambient 3hazy solution Water ambient <7 solids remained Water:acetone ambient 3hazy solution 1:2 Water:acetone ambient <3 solids remained 3:1 5.0MHCl:water ambient 7 clear solution 1:2 ^(a)Solubilities were calculatedbased on the total solvent used to give a solution; actual solubilitiesmay be greater because of the volume of the solvent portions used or aslow rate of dissolution. Values are rounded to whole number. Ifdissolution did not occur as determined by visual assessment, the valueis reported as “<”. If dissolution occurred as determined by the visualassessment after the addition of the first aliquot, the value isreported as “>”.

Small-Scale Scouting Experiments Towards the Development of ConditionsSuitable for Recrystallization

Approximately 16 small scale slurry experiments were carried out byvarying the slurry concentration, temperature, HCl acid molarconcentration, and content in the aqueous acetone mixtures as well asthe water content. Slurries of impure Form B were performed in a givensolvent system at targeted calculated concentration at ambient orelevated temperatures for various time/durations. The solids wereisolated by vacuum filtration and submitted for XRPD analysis. Thespecific experimental conditions are detailed in Table 12 where solventsystem ratios are by volume. The slurries in acidic aqueous acetonemixtures (Samples 1, 2, and 4) at ambient temperature failed to convertimpure Form B to pure Form B. FIG. 9 compares the XRPD patterns ofSamples 1 and 4 to the XRPD pattern of the starting material of theexperiments, impure Form B. FIG. 9 also compares Samples 1 and 4 to pureForm B material previously characterized in Example 6.

The slurries in acidic aqueous acetone mixtures at an elevatedtemperature of 50° C. (Samples 3 and 5) produced a disordered materialwith two broad low angle peaks material suggestive of a potentialmesophase. FIG. 10 compares XRPD patterns of Sample 3 and 5 to thestarting material of the experiments, impure Form B and to pure Form B.For comparison purposes, the samples were also compared to a secondimpure sample of Form B, (Impure Form B Sample 2 in FIG. 10 ). Thissecond impure Form B contained larger amounts of the unknown Form thanthe impure Form B previously described in Example 8. When increasing themolar concentration of HCl from 0.1 M (Sample 3) to 0.5 M (Sample 5),the intensity of these two peaks also increased.

Several slurry experiments were performed in water:acetone solventsystems starting with impure Form B and varying the water:acetone ratio,slurry concentration, and time. Based on the initial slurry results,experiments in 1:2 (v/v) water:acetone at ambient temperature wereperformed with aliquots taken after 16 hours (Sample 6) and 20.5 hours(Sample 7). The slurry in this solvent system was conducted at aconcentration of 100-125 mg/mL and ambient temperature. The XRPDpatterns of the resulting materials were consistent with pure Form B(FIG. 11 ). Using a water:acetone (1:2) solvent system resulted in a lowyield of 78-79% that was calculated for solids isolated by vacuumfiltration without drying.

In an effort to improve the yield, water:acetone 1:3 (v/v) was used at150 mg/mL concentration (Samples 13 and 14) however, the conversion wasnot completed even after 4 days (FIG. 12 ). Therefore, one experimentwas performed using water:acetone 1:2 (v/v) slurry for 18 hours and thenadding acetone to reach ratio water:acetone 1:4 (v/v) followed by slurryfor 4 hours (Sample 12). The XRPD pattern of the resulting material wasconsistent with Form B, however, one of the undesired peaks reappearedshifted from 3.95° 2θ to 4.2° 2θ (FIG. 12 ).

TABLE 12 Small-scale Slurry Experiment Conditions and Results Sample IDSolvent System^(a) Conditions Observation XRPD Result 1 0.5M HCl:acetone100 mg/mL Slurry, Dark yellow solids B + broad peaks at 1:1 RT, 3 days3.95 °2θ and 5.55 °2θ 2 2.0M HCl 150 mg/mL Slurry, Light yellow solidsB + A RT, 3 days 3 0.1M HCl:acetone 100 mg/mL Yellow orange Two lowangle 75:25 mobile suspension peaks in disordered material 4 0.1MHCl:acetone Slurry, 50° C., 15 h Bright yellow Form B + broad 1:2 mobilesuspension peak at 3.95°2θ 5 0.5M HCl:acetone 100 mg/mL Slurry, Darkorange mobile Two low angle 50:50 RT, 16 h suspension peaks indisordered material 6 water:acetone 1:2 100 mg/mL Slurry, Bright yellowForm B RT, 16 h mobile suspension 7 100 mg/mL Slurry, Bright yellow FormB RT, 20.5 h mobile suspension 8 100 mg/mL Slurry, Bright yellow Notanalyzed RT, 5 days mobile suspension 9 water:acetone 1:2 150 mg/mLSlurry, Form B + broad RT, 20 h smaller peak at 4.2 °2θ 10 150 mg/mLSlurry, Bright yellow Not analyzed RT, 4 days suspension with ribbon oforange solids below solvent line 11 water:acetone 1:2 125 mg/mL Slurry,Bright yellow Form B RT, 20 h mobile suspension 12 1. water:acetone1:2 1. 125 mg/mL, 1. Bright yellow Form B + small 2. water:acetone 1:4slurry, RT, 18 h mobile suspension broad peak at 2. Acetone added to 2.No observation 4.27 °2θ reach H2O:acetone 1:4 3. Bright yellow 3.Slurry, RT, 4 h mobile suspension 13 water:acetone 1:3 150 mg/mL Slurry,Form B + broad RT, 20 h peaks at 4.2 °2θ and 5.9 °2θ 14 150 mg/mLSlurry, Bright yellow Form B + broad RT, 4 days suspension with peaks at4.2 °2θ tiny ring of orange and 5.7 °2θ solids below solvent line 150.1M HCl:EtOH 100 mg/mL Slurry, Bright yellow Form B + broad 1:9 RT, 16h mobile suspension peak at 3.95°2θ 16 100 mg/mL Slurry, Bright yellowForm B + broad RT, 20.5 h mobile suspension peak at 3.95°2θ

Lara Controlled Laboratory Reactor Slurry Experiments

Several scale up experiments were carried out in efforts to demonstrateapplicable conditions for the conversion of impure Form B to pure FormB. The slurry conversion experiment was performed using a 1 Lround-bottomed controlled laboratory reactor (Radleys Lara CLR) equippedwith a Teflon anchor impeller, Julabo temperature control unit, andtemperature probe for monitoring of the reactor temperature throughoutthe experiment. The Julabo FP50 temperature control unit containedJulabo Thermal C10 fluid and the reactor temperature was measured with aK-type PTFE temperature probe. The experiments were carried out withLara Control software version 2.3.5.0. The software tracked circulatortemperature, vessel temperature, and stir rate, recording readings everytenth of a second throughout the experiment.

The reactor vessel was charged with the solids of impure Form B (58.86g) in 471 mL of a water:acetone 1:2 (v/v) solvent system achieving 125mg/mL slurry concentration (Samples 20-23). The resulting slurry wasstirred at 30° C. for up to 43 hours with stirring speed of 400 rpm. Theslurry was cooled to 25° C. over 30 minutes, discharged from the reactorvessel, and immediately slowly filtered (drop by drop) to dry land. Awater:acetone 1:2 (v/v) wash solution was prepared in advance and usedto wash the filter cake in one portion.

Pulls were taken usually at the 20th hour and if needed at later timepoints (Table 13). The scale up experiments showed that longer times andslightly elevated temperature (from ambient temperature to 30° C.) wereneeded at larger scale to convert impure Form B completely to pure FormB. Sample 22 was converted to pure Form B, while Samples 21 and 23 werenot analyzed. Sample 20 resulted in Form B, but a broad peak was alsoobserved at 4.2° 2θ.

TABLE 13 Scale-up Slurry Experimental Conditions and XRPD Results SampleSolvent System ID (v/v) Conditions XRPD Result 17 water:acetone 125mg/mL Slurry, B + broad peaks at 30 70 RT, 18 h 4.2 °2θ and 5.8 °2θ 18125 mg/mL Slurry, B + broad peak at RT, 23 h 4.2 °2θ 19 125 mg/mLSlurry, B + broad peaks at RT, 30 h 4.2 °2θ and 5.8 °2θ 20 water:acetone125 mg/mL Slurry, B + broad peak at 1:2 RT, 20 h 4.2 °2θ 21 125 mg/mLSlurry, Not analyzed RT, 45 h 22 125 mg/mL Slurry, Form B 30° C., 20 h23 125 mg/mL Slurry, Not analyzed 30° C., 20.5 h

TG-IR Characterization of Compound 2, Form B

The TG analyses were performed using a TA Instrument Q5000thermogravimetric analyzer. Temperature calibration was performed usingnickel and Alumel. The sample was placed in a platinum pan and insertedinto the TG furnace. The furnace was heated under a nitrogen purge to350° C. at a rate of 10° C./min.

Thermogravimetric infrared (TG-IR) analysis was performed on a TAInstruments Q5000 IR thermogravimetric (TG) analyzer interfaced to aMagna-IR 560® Fourier transform infrared (FT-IR) spectrophotometer(Thermo Nicolet) equipped with an Ever-Glo mid/far IR source, apotassium bromide (KBr) beamsplitter, and a mercury cadmium telluride(MCT-A) detector. The FT-IR wavelength verification was performed usingpolystyrene, and the TG calibration standards were nickel and Alumel™.The sample was placed in a platinum sample pan and the pan was insertedinto the TG furnace. The TG instrument was started first, immediatelyfollowed by the FT-IR instrument. The TG instrument was operated under aflow of helium at 90 and 10 cc/minute for the purge and balance,respectively. The furnace was heated under helium at a rate of 20°C./minute to a final temperature of approximately 140° C. IR spectrawere collected approximately every 32 seconds for approximately 7.5minutes. Each IR spectrum represents 32 co-added scans collected at aspectral resolution of 4 cm-1. Volatiles were identified from a searchof the High Resolution Nicolet Vapor Phase spectral library.

A TG-IR experiment was carried out on pure Form B (Sample 11 from thesmall-scale slurry experiments) at ambient temperature for 20 hours inan effort to investigate the stability of Form B at elevated temperatureby monitoring potential release of hydrogen chloride.

The TG data showed 6.4% weight loss at 33-137° C. (FIG. 13 ). Thecorrelation between the time and temperature is presented in Table 14.The series of IR spectra collected during the TG-IR experiment arepresented in FIG. 14 and FIG. 15 . The spectra demonstrated that onlywater was detected as a volatile and that no hydrogen chloride wasreleased.

TABLE 14 Correlation between Time and Temperature for TG-IR of Compound2, Form B Time (min) Temperature (° C.) Weight (%) 0.13 33.00 99.98 0.3836.60 99.93 0.55 40.20 99.87 0.73 43.80 99.77 0.90 47.40 99.63 1.0851.00 99.46 1.25 54.60 99.26 1.43 58.20 99.03 1.61 61.80 98.79 1.7865.40 98.52 1.96 69.00 98.22 2.14 72.60 97.86 2.32 76.20 97.44 2.5079.80 96.95 2.68 83.40 96.45 2.86 87.00 95.97 3.04 90.60 95.57 3.2394.20 95.23 3.41 97.80 94.95 3.59 101.40 94.71 3.77 105.00 94.50 3.95108.60 94.32 4.13 112.20 94.18 4.31 115.80 94.06 4.50 119.40 93.96 4.68123.00 93.86 4.86 126.60 93.78 5.04 130.20 93.71 5.22 133.80 93.65

Drying Experiments of Compound 2, Form B

Weighted amounts of impure Form B and pure Form B samples from theprevious experiments (Samples 14, 8, 11, 19, 21, and 23) were vacuumdried at ambient or elevated temperatures using various vacuum levelsfrom approximately 14 in Hg up 27-28 in Hg. The resulting materials wereweighted out prior to submission for XRPD analysis.

Two samples (Sample 14 and 8) were vacuum dried at 40° C. for 15 hours(approximately 29 in Hg) and demonstrated approximately 7.4% weightloss. One of the samples (Sample 8) was analyzed by XRPD and a newcrystalline XRPD pattern was obtained (FIG. 16 ) that was not consistentwith Form B.

Sample 11 was vacuum dried at ambient temperature for 0.5 hour(approximately 14 in Hg) demonstrating a 1.8% weight loss (calculatedfrom weighing the sample before and after drying). The XRPD pattern ofthe resulting material was consistent with Form B, however, tinyshifting in a few peak positions in the XRPD pattern was observed (FIG.17 ). Significant peak shifting was observed in the XRPD pattern ofSample 23 (FIG. 17 ) that was vacuum dried at ambient temperature for 1h (approximately 27-28 in Hg) demonstrating an 4.7% weight loss(calculated from weighing the sample before and after drying).

TABLE 15 Drying Conditions and Results for Compound 2, Form B WeightLoss (calculated by Sample weight before Source Conditions and afterdrying) XRPD Result Sample Vacuum oven, ~40° C., Weight loss: Notanalyzed 14 15 h (~29.5 in Hg) 7.5% Sample Vacuum oven, ~40° C., Weightloss: New crystalline 8 15 h (~29.5 in Hg) 7.4% material, not indexableSample Vacuum oven, 23° C., Weight loss: Not analyzed 14 2 h (~29.5 inHg) 6.8% Sample Vacuum oven, 22° C., Weight loss: Form B tiny peak 110.5 h (~14 in Hg) 1.8% shifting Sample Vacuum oven, RT, 1.0 h Weightloss: Form B + broad 19 (~27-28 in Hg) 9.7% peaks at 4.3 °2θ SampleVacuum oven, 22° C., Weight loss: B + broad peaks 21 0.5 h (~28 in Hg)30.6% at 4.2 °2θ and 5.8 °2θ Sample Vacuum oven, 22° C., Weight loss:Most likely Form 23 1.0 h (~27-28 in Hg) 4.7% B shifted

Conversion of Impure Form B to Pure Form B

The conversion of impure Form B to pure Form B was conducted inwater:acetone 1:2 (v/v) slurry at 125 mg/mL concentration and 30° C. for43 hours. Very slow filtration was observed and the wet cake wasair-dried at ambient conditions for 3.5 hours followed by vacuum dryingat ambient temperature and 15 in Hg for 0.5 hour and then at ˜27 in Hgfor 3.5 hours yielding 49.26 g (84%).

An XRPD pattern was obtained at different points in the conversion asshown in Table 16. After 42 hours of heating, XRPD analysis showed thatimpure Form B had completely converted to pure Form B. Once the materialwas filtered and dried, TG analysis was performed in addition to XRPDanalysis.

The XRPD pattern exhibited by the converted batch after drying wasconsistent with the XRPD pattern in FIG. 7 of pure Form B and its peaksaligned with the allowed peak positions from the pattern shown in FIG. 7. FIG. 18 compares the patterns of impure Form B, pure Form B ascharacterized in Example 6, and the pure Form B converted from impureForm B as described in Example 8.

The TGA data of the converted batch, Form B showed 7.5% weight lossbetween 31 and 120° C. (FIG. 19 ).

TABLE 16 Results of XRPD Analysis and TG Analysis during Conversion ofCompound 2 to Form B Analytical Point in Conversion Technique Result 125mg/mL Slurry, 30° C., 20 h XRPD Form B + small broad peak at 4.0 °2θ 125mg/mL Slurry, 30° C., 24 h XRPD B + broad peaks at 4.2 °2θ and 5.7 °2θ125 mg/mL Slurry, 30° C., 42 h XRPD Form B 125 mg/mL Slurry, 30° C., 43h XRPD Form B VF, washed with 50 mL of TGA 10% weight loss at 26-H₂O:acetone 1:2 Air-dried for 120° C. 3.5 h Vacuum drying, 22° C., 0.5h, XRPD Form B 15 in Hg Vacuum drying, 22° C. TGA 9.9% weight loss at26- 1.5 h, 27-28 in Hg 120° C. Vacuum drying, 22° C., 2.0 h, XRPD Form B27 in Hg TGA 7.5% weight loss at 31- 120° C.

We claim:
 1. A pharmaceutical composition comprising a pharmaceutically acceptable excipient for solid dosage deliver and between about 10 mg and about 1,000 mg of an isolated crystalline Form B of the di-HCl salt of structure:

characterized by an X-ray powder diffraction (XRPD) pattern comprising at least three 2θ values selected from 6.5±0.2°, 9.5±0.4°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.3±0.2°, 19.9±0.2°, and 22.4±0.2°. 